SpikeTides™ - Synthetic Proteotypic Peptides: light – heavy – quantified
SpikeTides™ represent fully synthetic customized peptides, which terminate as native C-terminal Arg or Lys peptides or consist of a tryptic fragment that is fused to a proprietary JPT-tag. The latter tagged peptides enable a cost efficient access to heavy labeled and/or quantified proteotypic peptides. JPT has developed a synthesis technology that enables ultra-fast, highly parallel and inexpensive synthesis of small scale peptides that are ideally suited for proteome wide profiling using SRM/MRM proteomic assays.
The sequences can be selected from bioinformatic prediction (i.e. Peptide Sieve) or from proteomic data repositories such as the Peptide Atlas (http://www.peptideatlas.org) that predict proteotypic peptides corresponding to peptides will be obtained from tryptic digestion of native proteomes.
Available SpikeTides Options:
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Small scale, unpurified proteotypic peptides with C-terminal Arg or Lys |
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Small scale, unpurified proteotypic peptides with tryptic C-terminal tag |
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Heavy labeled, small scale, unpurified proteotypic peptides with tryptic C-terminal tag |
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Quantified, small scale, unpurified proteotypic peptides with tryptic C-terminal tag |
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Quantified & heavy labeled , small scale, proteotypic peptides with tryptic C-terminal tag |
SpikeTides™
SpikeTides™ are customized, inexpensive, proteotypic peptides that terminate as C-terminal Arg/Lys.
Amounts: 50nmol/peptide
Purity: Unpurified
Application: Optimization and validation of multiplexed SRM assays
Price: from 14 US$ / 11 €
SpikeTides™_T
SpikeTides™_T are customized, inexpensive, small scale, unpurified proteotypic peptides with an C-terminal tag that can be cleaved by tryptic digestion.
Amounts: 50nmol/peptide
Purity: Unpurified
Application: Economic peptide source for multiplexed SRM assays
Price: From 12 US$ / 9 €
SpikeTides™_TL
SpikeTides™_TL are heavy labeled, proteotypic peptides that have a C-terminal tag that can be cleaved by tryptic digestion.
Amounts: 50nmol/peptide
Purity: Unpurified
Application: Development of SRM assays and relative quantification of proteins using a single product
Price: from 57 US$ / 36 €
SpikeTides™_TQ
SpikeTides™_TQ are quantified, proteotypic peptides that have a C-terminal tag that can be cleaved by tryptic digestion.
Amounts: 5 x 1 nmol/target peptide
Purity: Unpurified
Application : Absolute protein quantification
Price: from 68 US$ / 44 €
SpikeTides™_TQL
SpikeTides™_TQL are heavy labelled, quantified, proteotypic peptides that have a C-terminal tag that can be cleaved by tryptic digestion.
Amounts: 5 x 1 nmol/target peptide
Purity: Unpurified
Application: Absolute protein quantification
Price: from 105 US$ / 71 €
Benefits
- Thousands of SpikeTides™ for MRM assays
- Unmatched turnaround times (10 000 peptides per week!)
- Delivery in ready-to-use microtiter plates
- Aliquotation and mixing service available
- SpikeTides™ approach is more robust than other peptide assays and more cost effective than AAA
- Is fully compatible with downstream analyses like MS and HPLC
- Enables chemical barcoding of complex SpikeTides™ mixtures within the JPT-tag
Applications
- Optimization and validation of SRM assays
- Relative and absolute quantification of proteins
- Selection of best proteotypic peptides showing high mass spectrometry signal response
- Find predominant peptide fragments specific for your proteotypic peptide to be used in MRM transition
- Develop kits to quantify entire proteomic pathways
Background
Peptide quantification is a prerequisite in many areas of proteomics. One approach for unambiguous quantitative analysis of targeted proteins in complex mixtures is the use of tandem mass spectrometry to monitor one or more proteotypic peptide(s) from one protein of interest by a selected reaction monitoring (SRM) assay or by parallel analysis of many proteotypic peptides by a multiple reaction monitoring (MRM) assay. Absolute quantification is performed by the use of stable isotope-labeled proteotypic peptides as internal standards. These standards have to be purified to high level enabling subsequent AAA or alternative peptide quantification method (LavaPep, Ninhydrin, Lowry). This results in prices of several hundred US$ / € per peptide. JPT overcomes this situation by attaching a small chemical tag (which is cleavable by trypsin or other proteases) to the proteotypic peptide (proteotypic peptide + tag = SpikeTides™. Within the sample spiked with the appropriate SpikeTides™ (or mixture of SpikeTides™) the used protease (mostly trypsin) will cleave the peptide-tag bond releasing the desired proteotypic peptide and the JPT-tag in a 1:1 ratio.
Selected References:
High-throughput Generation of Selected Reaction-Monitoring Assays for Proteins and Proteomes
Picotti et al., Nature Methods (2009)
Full Dynamic Range Proteome. Analysis of S. cerevisiae by Targeted Proteomics
Picotti et al., Cell (2009)
More references under JPT Publications/Literature
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