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Frequently Asked Questions

Please refer to the peptide product categories below for frequently asked questions (FAQs). If you can not find the question you are looking for, please contact our customer support team: peptide@jpt.com

Ordering via our online Catalog (shop.jpt.com)

Do I need to create an account to place an online order for catalog products?

No, you can order without registrering. However ordering with an account gives you the advantage of having all your data saved in our system, including your adress and previous orders.

How do I create an account?

You can create a new account by clicking "new customer?" on the top right side of our webshop. Alternatively, can you create your new account directly in your cart before checkout.
The chosen password needs to contain at least 6 characters.

How can I find a product in JPT's catalog (online shop)?

There are several ways to find a brand or product in JPT's Webshop.
1. Choose from our list of applications, indications, protein names or organisms by clicking on the corresponding drop down menu of the Product Finder.
2. Type the product name or code, protein ID or any other keyword into the search box of the Product Finder.
3. In the main navigation menu at the top of every page, you can select the type of product you are looking for.

Do prices include sales taxes/VAT/MwSt.?

No, prices do not include tax. For shipment within Germany 19% MwSt will be included at checkout and in the invoice. For shipment within EU-countries you will need to provide the valid VAT number. International customers should note that all duties, import fees, and taxes are to be paid by the customer!

Are prices subject to change?

Product prices are subject to change occasionally and without warning. We apologize in advance for any inconvenience this may cause. However, prices shown on the day you order will be the invoiced prices.

Does JPT apply bulk discounts?

Yes, they are applied automatically at checkout.

Orders & Shipments

How much does JPT charge for shipping?

See details under our section "Shipping Prices",

How do I check the status of my order?

You will receive an acknowledgement of receipt via email directly after placing your order. We will then send an order confirmation as soon as the order has been processed (usually within 1 business day for catalog products, several weeks for customized products) and an email carrying all shipping information as soon as the order has been shipped.

Can I track my package online?

You will receive an email from our customer support when the order has been shipped. For shipments outside of the EU, this email also provides the Fedex tracking number and all needed order and shipping information.

When will my order arrive?

Orders will usually be processed within 24 hours. Most catalog products are ready to be shipped within 1 business day. Customized Products are ready within weeks. Please check the availability for each product within our Webshop. The additional carrier shipping time is listed below:
Shipment within Germany: 1 day
Shipment to EU countries :2 days
Shipment to the USA & Canada: 2-3 days
Shipment to other countries 3-4 days

Does JPT ship internationally?

Yes, we accept orders from and ship to all countries.

How do I return a product?

Return of products will be accepted only with prior notification via email (peptide@jpt.com)! Please contact us for return shipping instructions and to receive an authorization for return. Please send your unused/unopened return product, along with a copy of your original receipt to our head office.

What do I do if a product arrives damaged?

If there ever is a problem with your order, please contact us immediately (at peptide@jpt.com) for return shipping instructions and to receive an authorization for return. All damaged goods need to be returned to JPT's head office. Upon receipt a replacement will be sent.

How do I ask questions or make comments?

Please email your comment or question to peptide@jpt.com.

BioTides™

What is a BioTide™?

Biotides™ are the most inexpensive source of custom synthesized biotinylated peptides for screening application relying on the interaction between biotin and streptavidine.

How much peptide do I receive?

50 to 250 nmol per peptide.

How quickly can I get my BioTides™?

Up to 5000 BioTides™ will be delivered within 10 working days.

Where is the Biotin located and how is it attached?

The Biotin is located at the N-terminal end of the peptide and is attached via a flexible linker system enabling optimal interaction with the streptavidine.

How will BioTides™ be synthesized?

BioTides™ are synthesized on planar cellulose surfaces in a stepwise manner from the C- to N-terminus using preactivated Fmoc amino acids. After synthesis, BioTides™ are cleaved from the membrane. This synthetical method is called "SPOT synthesis" and guarantees synthesis of high numbers of peptides in very short times.

How is the C-terminal end of BioTides™ defined?

Standard BioTides™ consist of an additional glycine acid or amide at the C-terminus due to technical reasons. Individual C-terminal amino acids are also feasible at added costs (minimum order: 384 peptides).

What is the purity of BioTides™?

All BioTides™ are synthesized as non-purified materials. The purity of each BioTide™ depends on its primary sequence. Typically the biotinylated peptide represents the main product.

How long can a BioTide™ be?

Since purity of BioTides™ drops with increasing length we do not recommend BioTides longer than 20 amino acids.

How are BioTides™ QC’d?

5% of BioTides™ will be analyzed by LC-MS. In addition, JPT performs LC-MS of technical control sequences for each synthesis batch. Additional LC-MS analyses is available upon request.

What is the delivery format of BioTides™?

BioTides™ are delivered in ready-to-use in 96-well microplates (one freeze dried peptide per well, not immobilized, ready-to-dilute).

Is there a minimum order size?

Minimum order size is 24 peptides. For peptides with individual C-terminal amino acids minimum order size is 384 peptides.

Custom Peptide Synthesis

How does JPT Peptide Technologies QC its products?

With exception of our crude peptides, which will only be checked for identity by MALDI-MS, all synthetic peptides will be analyzed by HPLC and MS. Beside MALDI-MS our routine application involves HPLC-MS techniques to guarantee that a peak visible in the HPLC profile corresponds to the target mass. Since peptides show different ion response depending on the actual primary sequence, availability of different MS-equipment is crucial for doubtless analysis. At JPT Peptide Technologies’ state of the art equipment (MALDI-MS, HPLC-(ESI)MS (ion trap and quadrupole) guarantees unambiguous analysis. In addition, JPT Peptide Technologies offers optional amino acid analysis and peptide content determination. Analytical Data Sheets and analytical data are delivered with each peptide.

Is there any limitation on the length of peptides that can be achieved?

JPT Peptide Technologies offers custom service for peptides ranging from 2 to appr. 100 amino acids. Standard stepwise solid phase procedures provide access to most of the peptides in range from 5 to 50 amino acids in length. For short peptides (i.e. dimers, trimers etc.) and peptides with special modification we have solution phase approaches available. For long peptides up to appr. 100 amino acids we use our proprietary convergent peptide synthesis approach. For longer peptides the success rate drops below 50% and therefore recombinant technologies might be advantageous.

What purity levels are available and which purity is needed for my application?

JPT Peptide Technologies offers different purity levels starting with >70% up to >95%. Ultra pure materials with >97% purity are available upon request. The following list might serve as guideline for peptide specification:
>70%: immunological applications, epitope discovery, and nonsensitive screening;
>80%: immunological applications and epitope validation;
>90%: SAR studies, immune monitoring assays;
>95%: NMR, crystallization, enzymatic assays;
>97%: NMR, crystallization, sensitive bioassays.

At what scale are peptides synthesized?

JPT Peptide Technologies synthesizes custom peptides in a range from mg (smaller aliquots available) up to several grams per peptide.

How do I dissolve my peptides?

Solubility of peptides greatly depends on various parameters like primary and secondary structure, nature of modifications made, solvent, pH value and final concentration. Therefore, solubility is hardly to predict. If a peptide is difficult to solubilize in aqueous solution, sonification will support solubility. For basic peptides, 10% acetic acid has been recommended whereas for acidic peptides, aqueous ammonia or 10% ammonium bicarbonate solution were shown to be helpful. For peptides having poor solubility in aqueous solutions, organic solvents have to be applied. Use the minimal amount of organic solvent (DMSO is preferred but DMF, DMA, isopropanol, methanol and others can also be used) to dissolve your peptide. We strongly recommend to dissolve the peptide first in the organic solvent. In a second step, water or other aqueous media should be added in stepwise manner until the final concentration is reached.

How do I solubilize peptides that don't go into solution using aqeous buffers?

We do suggest suggest the following:

a) Start with pure DMSO to dissolve the peptides
b) Apply a small volume first and use vortexing and/or sonication
c) Apply more DMSO and vortex/sonicate again if peptides don't go in solution
d) You may consider higher temp. 40-50°C if points a-c don't work
e) Once the peptides are dissolved in pure DMSO start dilution with buffer
f) Important: use small volumes to dilute DMSO in a stepwise manner. Always add the buffer to the DMSO
g) Use sonication/vortexing after each addition of buffer
h) Watch out for a cloudy solution. This is a sign for of precipitation. Use sonication again to get rid of minor precipitation

Does JPT Peptide Technologies provide the modifications I need?

JPT Peptide Technologies provides a large variety of well established peptide modifications like acetylation, biotinylation, phosphorylation and attachment of fluorescent dyes or quencher pairs but is also specialized in doing unusal reactions. It will be a pleasure to discuss your special needs in detail.

What kinds of peptide endings are most appropriate for my studies?

The default ends of a peptide are a free amine group at the N-terminus and an acid group at the C-terminus. However, a peptide often represents a part of a parental protein sequence. In that case, the more “native” ends would relate to blocked N-terminus (i.e. acetyl) and C-terminus (i.e.amide). In addition, this modification avoids the introduction of additional charges and stabilizes the resulting peptide towards enzymatic degradation resulting from exopeptidases.

How many peptides can JPT Peptide Technologies synthesize?

JPT Peptide Technologies is known for its patented high throughput technologies. Up to one hundred thousand isolated peptides can be produced in weeks.

What kinds of synthesis methods are available?

JPT Peptide Technologies possesses a complete range of peptide synthesis techniques including: solid phase synthesis and solution phase synthesis techniques BOC chemistry and Fmoc chemistry stepwise synthesis, convergent synthesis approaches, ligation techniques, nano scale peptide synthesis to multigram scale manual peptide synthesis and automated proprietary high throughput peptide synthesis.

What is net weight and peptide content?

Peptides are normally delivered as fluffy lyophilized material. Due to the nature of the peptides, they may still contain traces of moisture and counter ions on protonated amino functions (N-terminus, Arg, His, Lys, etc.). This is not considered an impurity, but reduces the actual peptide content by approximately 10 to 30%. The net peptide weight is the actual weight of the peptide minus the additional weight from residual moisture and counter ions. In order to ensure an accurate peptide concentration, the non-peptide weigth needs to be considered and substracted from the gross peptide weight. An accurate determination of the peptide net weight can be performed by quantitative amino acid analysis, which is also available upon request.

How long does it take from order to delivery?

Routinely, assembly and purification and quality control of medium size peptides takes 3-4 weeks. In addition, JPT Peptide Technologies offers rush order options guaranteeing extremely short delivery times upon request.

How does JPT Peptide Technologies ship the peptides? Which data will be provided?

All peptides will be shipped lyophilized in small polypropylene vials (2 ml or 10ml) with screw caps. Alternatively, smaller aliquots will be shipped in small amber type glass tubes (1.8ml) with screw caps. All tubes contain labels with information on primary sequence, amount and batch number. The peptides provided will be accompanied by the original analytical data achieved and an analytical data sheet providing comprehensive information about key characteristics of the peptide such as primary sequence, molecular mass, morphology, counter-ions, purity, etc.

What are the best storage conditions and how stable are the peptides?

Generally, most peptides are stable over years when stored at -20°C in freeze dried state. However, peptides containing amino acids such as Met, Cys or N-terminal Gln tend to oxidation or pyroglutamate formation. Since kinetics of such deterioration depends on primary peptide structure, stability can hardly be predicted. Therefore, we recommend usage of peptides within 6 months.
Recommended storage conditions are: -20°C or colder in a dry environment. Avoid repeated freeze-thaw cycles. Avoid long-term storage of aliquots in solution. If solubilized (ex. in DMSO) aliquots can’t be avoided, store at -80°C.

Enzyme Profiling - Microarrays

Do you have a specific protocol used for the application of Enzyme Profiling microarrays?

JPT Peptide Technologies provides a detailed protocol together with every Enzyme Profiling microarray including slide layout and sequence list in a .gal-file format. The protocols can also be downloaded from our website.

Can the microarrays be used again?

JPT Peptide Technologies recommends using the microarrays only once.

How are the microarrays shipped and how should the microarrays be stored upon arrival?

The microarray slides are shipped at room temperature. For long-term storage, JPT Peptide Technologies recommends storage at +4°C.

What is the chemical stability of the microarrays / immobilized peptides?

As long as the microarrays are kept dry and cool (+4°C), the microarrays are stable for 18 months from date of delivery.

Do I need special reagents/chemicals to perform the assay?

For Kinase Profiling microarrays, the kinase in question, radioisotopic labeled ATP as well as assay buffer are needed. If fluorescence read-out should be performed, a suitable fluorescently labeled antibody or phosphospecifc stain is needed.
For Phosphatase Substrate Sets, the phosphatase in question and the appropriate assay buffer are needed. The read-out of the incorporated or cleaved phosphate is performed using anti-pTyr antibodies (JPT recommends Cell Signalling anti-pTyr-100 followed by Pierce anti-mouse-Dylight649).
For Protease Substrate Set, the protease in question and the appropriate assay buffer are needed. The read-out of the full-length or cleaved peptides is performed using anti-pTyr antibodies (JPT recommends Cell Signalling anti-pTyr-100 followed by Pierce anti-mouse-Dylight649)

What hardware is needed for the read-out of the Enzyme Substrate Sets?

For radioisotopic read-out of the kinase profiling microarrays, a system for detecting incorporated phosphate is needed. JPT recommends using a phosphoimaging screen. If a fluorescence read-out is performed, a microarray scanner with appropriate wave-length and laser settings is needed. For details, please refer to the protocol or contact our Customer support. For read-out of the phosphatase or protease profiling microarrays, a microarray scanner with appropriate wave-length and laser settings is needed. For details, please refer to the protocol or contact our Customer support.

Enzyme Profiling - Substrate Sets (Peptide Collections)

Do you have a specific protocol used for the application of Enzyme Profiling Substrate Sets?

JPT Peptide Technologies provides a detailed protocol together with every Enzyme Substrate Set including plate layout and sequence list in Excel-format. The protocols can also be downloaded from our website.

Are of the peptides' termini modified in any way?

The peptides used in the Kinase Substrate Sets are N-terminally biotinylated and C-terminally amidated.
The peptides used in the Phosphatase Substrate Sets are N-terminally free and C-terminally amidated.
The peptides used in the Protease Substrate Sets carry Fluorophore and Quencher on N- and C-terminal ends respectively.

How much enzyme is needed for the assay?

The enzyme concentration is mainly depending on the specific activity of your enzyme. If you already have experimental data on you enzyme please use these as orientation. If there is no information on enzyme activity available, we recommend to start with a high concentration and adjust later if necessary.

Can the plates be used again?

Peptide quantities are optimized for a single use.

Can I reorder the single substrates?

All kinase substrates are available in larger quantities and can be purchased off-the-shelf.
For phosphatase and protease substrates customized synthesis is necessary and can be ordered fast and economically. Please contact our customer support at peptide@jpt.com.

What is the chemical stability of the peptides?

Generally, most peptides are stable over years when stored at -20°C in freeze dried state. However, peptides containing amino acids such as Met, Cys or N-terminal Gln tend to oxidation or pyroglutamate formation. Since kinetics of such deterioration depends on primary peptide structure, stability can hardly be predicted. Therefore, we recommend usage of peptide sets within 6 months. Once thawed and dissolved in buffer solution, JPT recommends performing the assay without further delay to avoid degradation.

How are the plates shipped and how should the plates be stored upon arrival?

The plates are shipped freeze dried at room temperature. However, for long-term storage, JPT Peptide Technologies recommends storage at -20°C.

Do I need special reagents/chemicals to perform the assay?

For Kinase Substrate Sets, the kinase in question, radioisotopic labeled ATP, as well as assay buffer are needed. If fluorescence read-out should be performed, a suitable fluorescently labeled antibody or phosphospecific stain is needed.
For Phosphatase Substrate Sets, the phosphatase in question and the appropriate assay buffer are needed. As read-out reagent JPT peptide Technologies recommends "Biomol green". For more details, please refer to the protocol.
For Protease Substrate Set, the protease in question and the appropriate assay buffer are needed.

What hardware is needed for read-out of the Enzyme Substrate Sets?

For radioisotobic read-out of the kinase substrate set, a Streptavidin membrane (Promega, SAM-membrane) is needed, as well as a system for detecting incorporated radioisotopic labeled Phosphate (Phospho-Imager).
For the Phosphatase Substrate Sets, a microtiterplate-reader capable of detecting absorbance at 620-660nm is needed.
For the Protease Substrate Sets, a microtiterplate-reader capable of detecting fluorescence (Ex: 350nm, Em: 490nm). Preferred read-out procedure is from top but microtiterplates with clear bottom are also available.

Micro-Scale Peptides

What is a Micro-Scale Peptide?

Micro-Scale Peptides are Custom Peptides synthesized by the fastest and most effective synthesis technology currently on the market!

How expensive is a Micro-Scale Peptide?

Peptide price starts at 7 US$ / €.

How much material do I receive?

50 to 250 ug per peptide

How quickly can I get my Micro-Scale Peptides?

Up to 10 000 peptides will be delivered within 10 working days.

How will Micro-Scale Peptides be synthesized?

Micro-Scale Peptides are synthesized on planar cellulose surfaces in a stepwise manner from the C- to the N-terminus using preactivated Fmoc amino acids. This synthetical method is called "SPOT synthesis" and guarantees the synthesis of high numbers of peptides in very short times.

How is the C-terminal end of Micro-Scale Peptides defined?

Standard Micro-Scale Peptides consist of an additional glycine acid or amide at the C-terminus, due to technical reasons. However, individual C-terminal amino acids are also feasible at added costs (minimum order: 384 peptides).

What is the purity of Micro-Scale Peptides?

All Micro-Scale Peptides are synthesized as non-purified materials. The purity of each Micro-Scale Peptides depends on its primary sequence.

How long can a Micro-Scale Peptide be?

Since purity of Micro-Scale Peptides drops with increasing length we do not recommend Micro-Scale Peptides longer than 20 amino acids.

How are Micro-Scale Peptide QC’d?

3% of Micro-Scale Peptides will be analyzed by LC-MS. In addition, JPT performs LC-MS of technical control sequences for each synthesis batch. Additional LC-MS analyses requested by customer cost 26 € /$ per peptide.

What is the delivery format of Micro-Scale Peptides?

Micro-Scale Peptides are delivered ready-to-use in 96-well microplates (one freeze dried peptide per well, not immobilized, ready-to-dilute). Alternatively, peptide can be provided in 384 well plates, deep well plates of micronic rack tubes.

Is there a minimum order size?

Minimum order size is 24 peptides. For peptides with individual C-terminal amino acids, minimum order size is 384 peptides.

PepMix™

What is a Pepmix™?

PepMixes™ are peptide pools generated by chemical synthesis and pooling of individual 15meric peptides overlapping by 11 amino acids, spanning entire protein antigens or of selected MHC class-I or class-II-restricted epitopes.

How much peptide is in the vial?

Each vial contains 25ug of each of the individual peptides.

Which PepMixes are available as pre-made pools?

JPT offers a continuously growing number of peptide pools. Currently, we have appr. 95 pre-made pools available.

What do I do if my protein of interest is not available as PepMix™?

Simply send us your antigen sequence or Swiss Prot accession to receive a quote on your customized PepMix. Contact: peptide@jpt.com.

Are you also providing individual peptides and matrix pools for T cell epitope identification?

Since all pools are made from individual synthetic peptides, we offer provision of single peptides and matrix pools for custom PepMixes™ at only a little extra cost. Simply send us your antigen sequence or Swiss Prot accession # to receive a quote on your customized PepMix. Contact: peptide@jpt.com.

Are you offering Custom Pooling and Aliquotation service?

Yes - we recommend purchase of freeze dried pools and single peptide aliquots for enhanced stability. Our Pooling and Aliquotation protocols are validated and certified according to DIN ISO 9001:2008 and GCLP guidelines. We guarantee presence of all peptides, high accuracy of aliquotation and batch-to-batch reproducibility. Please ask our customer support for optimal design of your pools.

How many peptides are in a mix?

Depending on the length of an antigen, a mix can contain between 20 and several hundred peptides.

How do you ensure that the pool does not contain byproducts that cause inhibition of T-cell responses or give false positive responses?

Danger of false positive T-cell responses or inhibition of T-cell responses by toxic contaminants depend on peptide specification. Together with its clinical collaboration partners, JPT developed synthesis and purification protocols which avoid presence of contaminants causing false positive T-cell responses or inhibition of T-cell responses by toxic byproducts to a large extent. In addition, JPT established QC procedures ensuring a high level of batch to batch reproducibility. Positive control pools will be QC’s for chemical purity and integrity as well as biological performance. JPT’s pooling strategy is validated and was audited to be in accordance with DIN ISO 9001:2000 and GCLP requirements.

How will PepMixes™ be delivered?

PepMixes™ will be delivered freeze dried at room temperature.

How do I store my PepMix™?

Freeze dried PepMixes™ should be stored at -20°C for long term storage.

How stable is my PepMix™?

Since PepMixes contain freeze dried peptides, stability is determined by the stability of the single peptides in the mix. Generally, most peptides are stable over years when stored at -20°C in freeze dried state. However, peptides containing amino acids such as Met, Cys or N-terminal Gln tend to oxidation or pyroglutamate formation. Since kinetics of such deterioration depends on primary peptide structure, stability can hardly be predicted. Therefore, we recommend usage of peptide pools within 6 months. For clinical trials, we offer stability testing via our custom Pepmix program. Avoid repeated freeze/thaw cycles of dissolved PepMixes!

How do I reconstitute my PepMix™?

The best solvent to dissolve a PepMix is DMSO, followed by slow dilution with an appropriate buffer system. Vortexing and sonication may ease the dissolution process. Please refer to our downloadable PepMix-protocol for details. Please note that dissolved peptides have a limited stability. Therefore, we recommend immediate freezing of aliquots which are not used shortly after the dissolution process. JPT also offers freeze dried pool aliquots.

What is the maximum concentration of DMSO I can use in my T-cell assay?

DMSO is a cytotoxic reagent. Based on literature, we recommend a maximum DMSO concentration of 1% (v/v).

What peptide concentration do you recommend for stimulation?

The recommended concentration for stimulating PBMC's is 1-2 µg of each individual peptide/ml.

Can I stimulate whole blood samples?

Although successful stimulation of whole blood samples has been described, applied concentration of peptide pool may be significantly higher compared to PBMC (peripheric blood mononuclear cells) stimulation.

What pools do you recommend for standardisation and positive controls of T-cell assays such as ELISA and CFC?

Both of our CEF pools (standard and extended) have been extensively used as positive, antigen specific control for CD8+ T cell stimulation in ELISPOT, intracellular cytokine and CTL assays. For efficiency in vitro stimulation of antigen-specific CD4+ and CD8+ T cells, we have developed a series of CMV related PepMixes such as the pp65-pool, the IE-1 pool, the IE-2 pool and others.

PepSpots™

Do you have a specific protocol used for the application of PepSpot membranes?

JPT Peptide Technologies provides a detailed protocol together with every PepSpot peptide membrane.

How does JPT Peptide Technologies QC the synthesis of membrane-bound peptides?

In addition to the membrane bound PepSpot™ peptides ordered, JPT Peptide Technologies assembles control peptides on the same membrane, which are connected via a cleavable linker system to the cellulose surface. After cleavage of these peptides from the membrane, their quality will be checked by MALDI-MS. The control peptides include specific sequences which are difficult to synthesize. The purity of the peptides synthesized varies for each peptide and is dependent on the sequence and length. Our analyses have determined that peptide purity is typically >70% for average 6-15 mers (by HPLC and mass spectroscopy).

Which end of the peptides is coupled to the cellulose membrane?

The peptides are covalently linked to the cellulose support via the C-terminus. For sufficient flexibility for binding studies, a two ß-alanine residue spacer or a short PEG spacer is inserted between the cellulose and each PepSpot™ peptide.

When do you recommend acetylation of the N-terminus?

N-terminal acetylation is recommended for peptide scans, because peptides are more stable to degradation and the uncharged N-acetyl better represents the region in the native antigen than a charged NH3+-group. There is no extra charge for acetylation.

Do you recommend to substitute cystein residues by serine residues?

Yes, as cysteine containing peptides are sensitive to oxidation and cyclization. Therefore, the substitution of Cys by Ser increases the peptide quality. Secondly, unspecific interactions between cysteine residues and antibodies/proteins could occur which give false-positive results. Finally, Cysteine is very scarcely an essential residue in antibody epitopes.

How are the data from the PepSpot™ incubation evaluated?

Detections can be performed with a chemiluminescence substrate in combination with an imaging system or, if not available, with a standard X-ray film and film cassette. Detections via fluorescence read-out are not recommended.

Which enzyme-labeled antibodies are recommended for detection ?

We recommend the use of horseradish peroxidase(HPR)-conjugated antibodies, while other enzymes such as alkaline phosphatase (AP) may also be used. Please note: It has often been observed that AP binds to peptide spots depending on their sequences, whereas HRP in almost all cases did not interact with the peptides. Please also note: Do not use sodium azide as a preservative for buffers with peroxidase as it is an inhibitor of the enzyme.

Can unnatural or modified amino acids be used in the synthesis?

The spot synthesis technology is compatible with D and L forms of amino acids as well as unnatural amino acids. However, the chemistry requires that the amino acids have an Fmoc protection group at their N-terminus and acid labile protecting groups on their side chains (e.g. Pbf, OtBu, Trt, Boc, tBu, etc). Please note: Some unnatural amino acids may give poor coupling yields due to sterical hindrance.

Can the attached peptides be cyclized or biotinylated?

Yes, JPT Peptide Technologies also offers Cys-Cys-cyclization and biotinylation for PepSpots™.

What length of offset do I need for peptide scans?

Most linear peptide epitopes for antibody applications are between 3-9 amino acids in length. To be on the save side, we recommend a peptide length of 13 amino acids and an offset of 2 to 3 amino acids for the mapping of linear epitopes. Hence, when working with a large protein, it is recommended that the offset does not exceed 3 amino acids in order to localize the epitope. An offset of 1 is recommended for finer mapping of the epitope. For the mapping of conformation dependent (discontinuous) epitopes, we recommend a peptide length of 15 to 20 amino acids and an offset 5 amino acids in maximum.

How much peptide is synthesized per Spot?

The amount of peptide that is synthesized is determined theoretically from calculating the amount of free amines available for coupling per Spot and assuming a coupling efficiency of >98% per cycle. A typical synthesis is expected to yield between 5-10 nmol (6-12 µg for an average 10mer peptide).

What is the size of a PepSpots™ peptide?

The PepSpots™ peptides have a diameter of approx. 2-3 mm.

What is the density of peptides on the membrane?

The peptides are synthesized in a grid fashion with 0.37 cm x 0.37 cm (0.15 in x 0.15 in, center-to-center) spacing, 20 peptides per row. Other formats are available upon request.

What is the chemical stability of the attached peptides?

JPT Peptide Technologies offers two types of PepSpots™ peptide membranes:
- PepSpots™: The connection between the peptide and the membrane is an ester bond and therefore labile towards strong bases (i.e. NaOH) and nucleophiles (i.e. NH2R). The peptides themselves are quite stable. However, strong acids may occasionally break acid labile amide bonds such as Asp-Pro. In addition, there are several amino acids such as Cys or Met which are sensitive towards oxidation. Finally, very strong bases (NaOH) may hydrolize peptide bonds as well.
- PepSpots™ (PEG): The connection between the peptide and the membrane is an ether bond and therefore stabile towards strong bases (i.e. NaOH) and nucleophiles (i.e. NH2R). We suggest to use this membrane type if electroblotting experiments or application of alkaline phosphatase is planned.

What is the chemical stability of the membranes?

The membrane material is stable to synthesis conditions, but should not be exposed to strong acids for prolonged periods.

What are the appropriate storage conditions for the membrane?

New membranes should be stored at -20°C until use. Incubated membranes which will be reused within several days should be kept with a small volume of T-TBS buffer in a Petri dish at 4°C. Incubated membranes which will be stored for a longer period should be regenerated and kept at -20°C.

Peptide Microarrays

What kind of hardware do I need to read-out my peptide microarrays?

You need a microarray scanner with the following specifications:
For low densitiy peptide microarrays (up to 3 x 384 Peptides/Microarray):
- Minimum scanning-resolution: 50 um / Pixel - Focusing should be possible - Intensity should be adjustable
- Size of slide: 1 x 3 inch
- Size of scanned area should be at least 63 x 21mm
- Extinction/emission filters depending on the fluophores used
For high densitiy peptide microarrays (more than 3 x 384 Peptides/Microarray):
- Minimum scanning resolution: 10 um / Pixel
- Focusing should be possible - Intensity should be adjustable - Size of slide: 1 x 3 inch
- Size of scanned area should be at least 63 x 21mm
- Extinction/emission filters depending on the fluophores used
- Software capable of reading and analyzing .gal-files.
JPT is working with a Genepix Axon 4000B, Genepix 4200 and a Genepix 4300 Scanner system. The resulting image should be an 8- or 16-bit format to be compatible with most microarray processing softwares.

Do you have specific protocols for peptide microarrays?

JPT Peptide Technologies provides a detailed protocol together with every peptide microarray. These can also be downloaded from our website.

How does JPT QC the production of peptide microarrays?

With each synthesis batch we run well selected control peptides that are analyzed by HPLC-MS and need to pass quality control. These control peptides include sequences that are difficult to synthesize. After printing each microarray slide is scanned using our high throughput-scanning device and the immobilized peptide array is checked for completeness and uniformity.

How are the peptides immobilized onto the microarrays?

Peptides are covalently linked to the microarray surface chemoselectively via the N-terminus. A flexible linker is inserted between peptide and microarray surface ensuring availablity of the binding sites within the peptide. By capping steps during synthesis we achieve that only full length peptides carry the N-terminal coupling moiety, thus enabling a high-throughput purification of all peptides during immobilization.

Do you recommend substitution of cystein residues by serine residues?

Yes, as cysteine containing peptides are very sensitive to oxidation and cyclization, substitution of Cys by Ser increases peptide stability. Additionally, unspecific interactions between cysteine residues and antibodies/proteins sometimes occur and give false-positive results. Finally, Cysteine is very scarcely an essential residue in antibody epitopes.

How are incubation results evaluated?

JPT recommends using fluorescence readout-systems in combination with dye-labeled antibodies. Since the glass surface is optimized for low background in fluorescence scanner systems, high S/N ratios can be achieved using high resolution laser scanners.

What is the concentration of each peptide on the microarrays?

The amount of peptide immobilized in one spot (100µm diameter) depends on the surface loading of the peptide microarray. Experiments showed a loading capacity of 4pmol/mm³. Therefore, the amount of peptide immobilized in one spot is approx 30 fmol/ peptide.

What is the size of a peptide microarray?

JPT´s peptide microarrays are compatible with most scanner system accepting microarrays of 1 x 3 inches (or 2.5 x 7.5 cm).

What is the size of the peptide spots on the peptide microarray?

The diameter of the individual spots is approx 80-140µm. The size mainly depends on the physicochemical properties of the peptide solution spotted and is very hard to predict.

What is the layout of JPT´s microarrays?

All our microarrays display each peptide in several copies to ensure verifiable results. The exact layout for each microarray is noted in the product description.

What is the chemical stability of JPT´s peptide microarrays?

The bond between peptides and microarray surface is stable in a range of pH 4 -10. Higher or lower pH values might lead to signal loss due to degradation.

What are the appropriate storage conditions for the microarrays?

Peptide microarrays should be stored at 4°C. JPT recommends NOT to freeze the microarrays since the effects of freeze thaw cycles on the glass surface are unclear. Properly stored, our peptide microarrays are stable for 18 months.

Is it possible to regenerate a peptide microarray?

JPT does not recommend regeneration of peptide microarrays by stripping bound proteins or antibodies from the surface because the surface can be damaged and also very low remainders of protein or antibody can cause a very high background.

PepStar™ Peptide Microarrays - Detection

Which amount of antibody is needed?

We need about 10-50 µg antibody per microarray experiment. Please inform us about buffer composition.

Is there a specific concentration you would like the antibody solution?

We would like to have the 10-50 µg of antibody as highly concentrated as possible.

What is the address to send the antibody solution?

JPT Peptide Technologies GmbH
Customer Support
Volmerstr. 5 (UTZ)
12489 Berlin, Germany
Always contact us for approval before you ship any product to us.

How should I send the antibody solution?

Do not ship any product to JPT without clearing the shipment procedure with our customer support (peptide@jpt.com)
The procedures for clearing antibodies/proteins through Customs have been tightened in 2011.
Therefore, please follow the recommendations below to avoid any problems by importing your goods.

1) Send us a list of the products to be sent and we will prepare a proforma invoice for your shipment

2) Please use FedEx or World Courier to ship your antibodies/proteins and provide a FedEx (or World Courier-) tracking number as soon as available. We will track the shipment and contact the carrier as soon as the package arrives in Germany to speed up the customs procedure.

3) In case of dry ice shipment please add enough dry ice for at least one week.

Peptide Conjugates

Why should my peptide or hapten be cross-linked to a carrier protein?

Molecules of small molecular weights such as peptides and haptens are insufficient immunogens when injected alone. These types of molecules should always be conjugated to a carrier protein when used for antibody production.

How do I select a carrier molecule for my peptide?

The carrier molecule you select depend on the species under examination. KLH is routinely used for vertebrate research because there are no homologous vertebrate proteins resulting in little if any non-specific antibody activity. For invertebrate studies, it is best to use a carrier such as BSA or ovalbumin.

Why is my conjugated peptide solution cloudy?

Depending on the structure and solubility of the carrier used, peptide conjugates have often limited solubility in water. However, this does not affect its immunogenicity and cloudy solutions can be used directly for immunizations.

How will my conjugated peptide be shipped?

Conjugates will be shipped as chilled solutions since it has been shown that both frozen solutions and freeze dried materials show sub-optimal immunization after reconstitution.

PepTrack™ Peptide Librairies

Which PepTrack™ option is best for my application?

For screening and discovery approaches, one of the most economical options Fast Track or Research Track might be best suited. Depending on the goal of the experiments, we may recommend a combination with full analytical coverage by LC-MS and/or an increased purity level (i.e. main product = target peptide). For all immune monitoring applications and assessment of vaccination trials, we recommend PepTrack™ (Discovery Track & Trial Track) options with guaranteed purity levels and full analytical coverage. Please contact our customer support for more details at peptide@jpt.com.

Why are JPT’s peptide libraries superior to those of other peptide vendors?

JPT maintains an DIN ISO 9001: 2008 certified Quality Management System since many years and was successfully audited to fully comply with Good Clinical Laboratory Practices (GCLP) for its custom peptide services to be used in clinical T-cell assays. Together with its governmental-, NGO- and industrial partners, JPT designs peptide synthesis and purification protocols that avoid formation and presence of false positive failure sequences and contaminants in ELISPOT, ICS and other T-cell assays. In addition, JPT ensures that no toxic contaminants will be present in its peptide preparation. Stringent documentation avoids presence of epitopes from foreign antigens and cross contaminations.
JPT provides exact purity information for all peptides that are analyzed. QC is performed by QUANTITATIVE LC-MS and NOT by QUALITATIVE MALDI-MS. JPT offers QUANTITATIVE QC on all peptides that are synthesized. JPT does not offer peptide purities based on misleading terms such as "average purity". All peptides will be produced at JPT’s main laboratories in Germany. JPT does not sell or distribute peptides from any other vendor. JPT’s raw material vendors are audited on a regular basis for reliability. Final peptides can be traced back to the level of raw incredients. Related documents are archieved for 10 years.

How do I solubilize my peptide library?

The solubility of peptides greatly depends on its primary and secondary structure and on the nature of modifications made. Many peptides are soluble in aqueous solution, basic peptides in 10% acetic acid and and acidic peptides in aqueous ammonia or 10% ammonium bicarbonate solution. Unfortunately, final solubility is hardly to predict. Therefore, for PepTrack™ libraries we suggest to apply DMSO as an organic solvent which enables complete dissolution of most peptides. In order to dissolve your peptides, apply the minimal amount of DMSO to dissolve your peptide and slowly add aqueous buffer or medium until the final concentration is reached. Please note that DMSO can oxidize peptides upon storage and has cell toxic properties when used in concentrations at elevated concentrations.

What are the best storage conditions and how stable are the peptides?

Peptides will generally be provided lyophilized as white fluffy materials which has been proven to be the best choice to avoid premature decomposition.
Ideal storage conditions are:
-storage at –20°C or colder in a dry environment.
-avoidance of repeated freeze-thaw-cycles. -try to avoid storage of aliquotes in solution, instead:
ORDER LYOPHILZED ALIQUOTES
-if storage in solution can not be avoided, store at –20°C and use sterile buffers ad slightly acidic pH.

How are Peptrack™ libraries delivered?

Fast Track: freeze dried in 96 well microplates with rubber mat
Research Track: freeze dried in 96-vial micronic racks (96 individual tubes) closed with individual rubber caps
Discovery Track: freeze dried in 96-vial micronic racks (96 individual tubes) closed with individual rubber caps or 4ml plastic tubes with screw cap
Trial Track: freeze dried in 96-vial micronic racks (96 individual tubes) closed with individual rubber caps or 4ml plastic tubes with screw cap

SpikeTides™

Which types of SpikeTides™ are available?

SpikeTides™: proteotypic peptides with c-terminal Lys or Arg residue.
SpikeTides™ T: proteotypic peptides with additional c-terminal tag. SpikeTides T need to undergo tryptic digestion to release the native proteotypic peptide.
SpikeTides™ TL: SpikeTides™ T labeled with stable isotopes carrying an additional c-terminal tryptic tag. SpikeTides TL need to undergo tryptic digestion to release the heavy labeled native proteotypic peptide.
SpikeTides™ TQ: SpikeTides T provided with information on absolute quantity for each peptide. SpikeTides™ TQ need to undergo tryptic digestion to release respective proteotypic peptide.
SpikeTides™ TQL: SpikeTides™ T fully quantified and labeled with stable isotopes carrying an additional c-terminal tryptic tag. SpikeTides™ TQL need to undergo tryptic digestion to release respective proteotypic peptide.

Which type of SpikeTides™ is the best one for my application?

SpikeTides™ are light proteotypic peptides intended for optimization and validation of multiplexed SRM assays.
SpikeTides™ L are isotopically labeled proteotypic peptides with C-terminal heavy Arg or Lys for development of SRM assays and relative quantification of proteins using a single product.
SpikeTides™ TQ are light and quantified proteotypic peptides that can be used for quantification of isotopically labeled proteins in biological samples (e.g SILAC).
SpikeTides™ TQL are isotopically labeled and absolutely quantified proteotypic peptides with C-terminal heavy Arg or Lys for absolute quantification of proteins in biological samples.

Why is the tag necessary?

The tag is necessary for realizing the peptide synthesis for heavy labeled peptides. In addition, the tag carries a moiety enabling the quantification of the target peptide during HPLC-runs using specific wavelengths. The tag is optimized to enable efficient cleavage using standard proteases like trypsin. After cleavage, the proteotypic peptide with the native C-terminus is available for the MRM assay.

Do you have a specific protocol used for the application of SpikeTides™ with or without tag?

JPT Peptide Technologies provides a detailed protocol together with every SpikeTides™ library including plate layout and sequence list in Excel-format.

Are the termini of the peptides modified in any way?

As necessary for proteotypic peptides, the N-termini are free, the C-termini are acidic.

What amount of peptide is in each well?

For the SpikeTides™, SpikeTides™ T and SpikeTides™ TL, a rough estimation of the peptide amount is given in the product documentation. The estimation of peptide amount is based on the determination of the amount of coupled amino acid in the last step. For the quantified SpikeTides™ TQ and SpikeTides™ TQL, the synthesized peptides is aliquoted in 5x1nmol fractions. Quantification is based on HPLC-MS analytics and UV read-out of the Quantification-tag.

What information will be provided?

An Excel-Spreadsheet will be provided giving all information concerning peptide sequence, sequence ID (if given), synthesis batch number and corresponding well in the microtiterplate/96 tubes rack.

How much enzyme is needed for the cleavage of the tagged SpikeTides™?

The cleavage of the SpikeTides™ can be performed in the same assay solution as the tryptic digest of the target protein takes place.

How do I know that the cleavage of peptide / tag was succesfull?

JPT Peptide Technologies validated the cleavage efficency for many different peptide surroundings. The efficiency of the cleavage can be checked by doing HPLC-runs after the cleavage was performed. The signal of the cleaved tag should bevisible at a retention time of 0 to 0.5min (C18, 3µ, 20x2mm, 1ml/min flowrate, 6 minute gradient, 5% to 95% Acetonitril water) and should light up when analyzed at 350nm. Uncleaved peptide /tag will have a signal at higher retention times (for examples and details about these validation steps, please contact our customer support at peptide@jpt.com).

What is the chemical stability of the peptides?

As long as the peptides are freeze dried and kept at low temperature, the stability is very good. Once thawed and dissolved in buffer solution, JPT Peptide Technologies recommends to perform the assay without further delay to avoid degradation of the peptides.

How are the plates shipped and how should the plates be stored upon arrival?

The plates or micronic racks are shipped at room temperature. For long-term storage JPT Peptide Technologies recommends to keep the plates /racks sealed at 4°C.

Do I need special reagents/chemicals to perform the assay?

A sample of protease trypsin is required to cleave the tag. Potentially, other required chemicals are described in the protocol and are dependent on the modifications of the library ordered.

Is there any other modification I need to consider in respect to my desired peptide?

JPT Peptide Technologies recommends to exchange all Cysteine residues by alkylated Cysteine and all Methione residues by sulfoxided Methionine.

What other delivery options are there?

JPT Peptide Technologies will be able to ship the peptides in different formats, plates and tubes. Default shipping formats are freeze dried peptides in sealed 96 wellplates (SpikeTides™, SpikeTides™ T and SpikeTides™ TL) or freeze dried aliquoted peptides in sealed 96 tube micronic racks (SpikeTides™ TQ and SpikeTides™ TQL). Both plates or racks are filled row-wise from the upper left corner. Changes in shipping formats can be discussed and evaluated with our customer support but might be subject to additional pricing.

Single Peptides

How will Single Peptides be delivered?

Peptides are delivered freeze dried at room temperature.

How does JPT Peptide Technologies QC its products?

All synthetic peptides are analyzed by HPLC and MS. Besides MALDI-MS, our routine application involves HPLC-MS techniques to guarantee that a peak visible in the HPLC profile corresponds to the target mass. Since peptides show different ion response depending on the actual primary sequence, availability of different MS-equipment is crucial for doubtless analysis. JPT Peptide Technologies' state of the art equipment (MALDI-MS, HPLC-(ESI)MS (ion trap and quadrupole) guarantees unambiguous analysis.

How do I dissolve my peptides?

Solubility of peptides greatly depends on various parameters like primary and secondary structure, nature of modifications made, solvent, pH value and final concentration. Therefore, solubility is hard to predict. If a peptide is difficult to solubilize in aqueous solution, sonification will support solubility. For basic peptides, 10% acetic acid has been recommended whereas for acidic peptides, aqueous ammonia or 10% ammonium bicarbonate solutions were shown to be helpful. For peptides having poor solubility in aqueous solutions, organic solvents have to be applied. Use the minimal amount of organic solvent (DMSO is preferred but DMF, DMA, isopropanol, methanol and others can also be used) to dissolve your peptide. We strongly recommend dissolving the peptide first in the organic solvent. In a second step, water or other aqueous media should be added in stepwise manner until the final concentration is reached.

What are the best storage conditions and how stable are the peptides?

Generally, most peptides are stable over years when stored at -20°C in freeze dried state. However, peptides containing amino acids such as Met, Cys or N-terminal Gln tend to oxidation or pyroglutamate formation. Since kinetics of such deterioration depends on primary peptide structure, stability can hardly be predicted. Therefore, we recommend usage of peptides within 6 months. Recommended storage conditions are: -20°C or colder in a dry environment. Avoid repeated freeze-thaw cycles. Avoid long-term storage of aliquots in solution. If solubilized aliquots can´t be avoided, store at -20°C and use sterile buffers at slightly acidic pH.

What to do if my peptide of interest is not available in JPT´s list?

Simply send us your peptide sequence, needed amount and purity to receive a quote on your custom peptide. Contact: peptide@jpt.com

JPT 3D-Epoxy Glass Slides

How are JPT 3D-Epoxy Glass Slides delivered?

Slides will be shipped chilled on dry ice and are sealed under an inert atmosphere (Argon).

How many slides are sealed in one box?

We can offer delivery of slides in sealed boxes containing 5 or 25 slides.

If I did not use all slides in the box, should I reseal it under argon?

We do not recommend resealing of an already opened box since moisture may have condensed immediately after opening at the slide surface that can't be removed by resealing.

How stable are the slides?

For slides in ORIGINALLY sealed boxes, the expiration time is 6 months.
For slides in opened boxes, we are not able to give definite data since stability of the reactive slides very much depends on the environmental conditions such as humidity and temperature. We suggest using such slides within a few days.

How do I store my JPT 3D-Epoxy Glass Slides?

We recommend storing the sealed slides at 4 to 10°C. Make sure to leave the sealed slides at room temepratur to allow them to adjust well before usage! Please avoid taking cold slides out of the sealed boxes since moisture would condensate rapidly on the surface thus leading to a reduced reactivity.
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