Phosphorylation Site Detector

JPT Peptide Technologies offers an efficient way to detect potential phosphorylation sites in kinase substrate proteins. Each Phosphorylation Site Detector represents a single peptide microarray on a glass slide containing an overlapping peptide scan through known proteins (please refer to table below). Subsequent to incubation with target kinase in the presence of radioactive ATP, incorporated phosphate can be detected by autoradiography or phosphorimaging. Alternatively, phospho-specific antibody-based fluorescence readout is applicable for phosphopeptide detection.

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Benefits of Phosphorylation Site Detector microarrays

Save time and material to determine phosphosites in proteins
Screen several hundred peptides economically
Learn more about your kinase/kinase target protein

Applications for Phosphorylation Site Detector microarrays

Identification of kinase substrate peptides
Detection of potential phosphosites in substrate proteins
Determination of auto-phosphorylation sites
Identification of upstream kinases for your target protein
Elucidation of signal transduction pathways

Available Phosphorylation Site Detector microarrays

Protein	Product-Code	Organism	Primary accession number	Database	No. of Peptides	Scan type*
14-3-3 zeta	PD-001	homo sapiens	NP_003397	Swiss-Prot	79	13/10
AKT1	PD-002	homo sapiens	NP_005154	NCBI	157	13/10
BCL 2	PD-003	homo sapiens	NP_004313	NCBI	79	13/11
B-Myb	PD-005	homo sapiens	NP_002457	NCBI	345	13/11
CARS	PD-061	homo sapiens	Q13427	Swiss-Prot	149	15/10
CD5 antigene	PD-006	homo sapiens	NP_055022	NCBI	41	13/11
Cdc25C	PD-007	homo sapiens	NP_001781	NCBI	155	13/10
CHK2	PD-008	homo sapiens	NP_009125	NCBI	178	13/10
c-jun	PD-009	homo sapiens	NP_002219	NCBI	160	13/11
Connexin 43	PD-011	homo sapiens	NP_000156	NCBI	94	13/9
crystallin B	PD-012	homo sapiens	NP_001876	NCBI	82	13/9
Cyp11	PD-013	homo sapiens	P30405	Swiss-Prot	195	13/10
Cyp18.1	PD-014	homo sapiens	Q9H2H8	TrEMBL	149	13/10
Cyp18.2	PD-015	homo sapiens	Q9Y536	TrEMBL	152	13/10
Cyp18.6	PD-016	homo sapiens	Q9BXZ1	TrEMBL	153	13/10
Cyp19.2	PD-017	homo sapiens	O43447	Swiss-Prot	165	13/10
Cyp23	PD-018	homo sapiens	P23284	Swiss-Prot	196	13/10
Cyp23a	PD-062	homo sapiens	P45877	Swiss-Prot	67	15/12
Cyp33	PD-063	homo sapiens	Q9UNP9	Swiss-Prot	97	15/12
Cyp40	PD-064	homo sapiens	Q08752	Swiss-Prot	120	15/12
Cyp57	PD-065	homo sapiens	Q8WUA2	Swiss-Prot	160	15/12
Cyp60	PD-066	homo sapiens	Q13356	Swiss-Prot	170	15/12
Cyp73	PD-067	homo sapiens	Q96BP3	Swiss-Prot	212	15/12
CYPA	PD-019	homo sapiens	P62937	Swiss-Prot	77	13/10
CypL1	PD-068	homo sapiens	Q9Y3C6	Swiss-Prot	52	15/12
CypL6	PD-069	homo sapiens	Q8IXY8	Swiss-Prot	100	15/12
E2F	PD-020	homo sapiens	NP_005216	NCBI	143	13/10
EpoR	PD-021	homo sapiens	NP_000112	NCBI	75	13/10
FKBB12.6	PD-022	homo sapiens	P68106	Swiss-Prot	48	15/13
FKBP12	PD-023	homo sapiens	P62942	Swiss-Prot	48	15/12
FKBP13	PD-024	homo sapiens	P26885	Swiss-Prot	66	15/12
FKBP19	PD-026	homo sapiens	Q9NYL4	Swiss-Prot	95	13/11
FKBP22	PD-027	homo sapiens	Q9NWM8	Swiss-Prot	100	15/12
FKBP23	PD-028	homo sapiens	Q9Y680	Swiss-Prot	124	15/12
FKBP25	PD-029	homo sapiens	Q00688	Swiss-Prot	107	15/12
FKBP36	PD-030	homo sapiens	O75344	Swiss-Prot	106	15/11
FKBP37	PD-031	homo sapiens	O00170	Swiss-Prot	107	15/11
FKBP38	PD-032	homo sapiens	Q14318	Swiss-Prot	115	15/11
FKBP44	PD-033	homo sapiens	Q9NZN9	Swiss-Prot	125	15/11
FKBP51	PD-034	homo sapiens	Q13451	Swiss-Prot	149	15/11
FKBP52	PD-035	homo sapiens	Q02790	Swiss-Prot	150	15/11
FKBP63	PD-070	homo sapiens	O95302	Swiss-Prot	140	15/11
FKBP65	PD-071	homo sapiens	Q96AY3	Swiss-Prot	144	15/11
GAB1	PD-036	homo sapiens	NP_002030	NCBI	342	13/11
LKB1	PD-038	homo sapiens	NP_000446	NCBI	141	13/10
MDM2	PD-040	homo sapiens	NP_002383	NCBI	161	13/10
MeCP2	PD-041	homo sapiens	NP_004983	NCBI	159	13/10
NFATP	PD-043	homo sapiens	NP_775114	NCBI	305	13/10
NKTK	PD-072	homo sapiens	P30414	Swiss-Prot	208	15/8
Nucleophosmin 1	PD-044	homo sapiens	NP_002511	NCBI	95	13/10
p53	PD-045	homo sapiens	NP_000537	NCBI	96	13/9
PAK1	PD-046	homo sapiens	AAC24716	NCBI	181	13/10
Par17	PD-073	homo sapiens	Q9Y237-2	Swiss-Prot	48	15/12
Parvulin14	PD-047	homo sapiens	Q9Y237	Swiss-Prot	60	13/10
PDK1	PD-048	homo sapiens	NP_002604	NCBI	182	13/10
Phas-I	PD-049	homo sapiens	NP_004086	NCBI	54	13/11
Pin 1	PD-050	homo sapiens	Q13526	Swiss-Prot	76	13/11
PTEN	PD-052	homo sapiens	NP_000305	NCBI	196	13/11
PTMA	PD-053	homo sapiens	NP_002814	NCBI	50	13/11
PTPA	PD-074	homo sapiens	Q15257-2	Swiss-Prot	78	15/11
PTPN1	PD-054	homo sapiens	NP_002818	NCBI	142	13/10
RAD9	PD-055	homo sapiens	NP_004575	NCBI	96	13/9
RAF1	PD-056	homo sapiens	NP_002871	NCBI	319	13/11
RBL2	PD-057	homo sapiens	NP_005602	NCBI	377	13/10
S6 protein	PD-058	homo sapiens	NP_001001	NCBI	80	13/10
Tie 2	PD-060	homo sapiens	Q02763	Swiss-Prot	172	13/11

* The first number X indicates the total number of amino acids of the peptide-sequence, whereas the second number Y indicates the number of overlapping amino acids.

Figure:
For quality control, four identical subarrays (including controls) are printed to each single microarray. All peptides are chemoselectively immobilized via their N-terminus. The hydrophilic linker moiety 1-amino-4,7,10-trioxa-13-tridecanamine succinimic acid is used as a spacer between the peptides and the glass surface.

Figure:
The full-length control proteins are covalently attached by non-selective immobilisation chemistry using the amino-function of the lysine-side chains. We can not guarantee that the proteins are natively folded.

	Protein	Company	Cat#
VIII	Tau-Protein	Sigma	T-9392
VII	β-Casein	Sigma	C-6905
VI	α-Casein	Sigma	C-7891
V	MBP	Biomol- Upstate	13-104
IV	Histon 4	Biomol- Upstate	14-412
III	Histon 3	Biomol- Upstate	14-411
II	Histon 2A	Biomol- Upstate	14-409
I	Histon 1	Biomol- Upstate	14-155

Testimonials for Phosphorylation Site Detector:

"We have successfully applied JPT's peptide microarrays as a screening tool for potential kinases transfering phosphate to phosphorylation sites identified by mass spectrometry. The triplicate alignment as well as the thorough selection of controls enabled generation of high quality data confirming our in vivo data generated by means of affinity chromatographic procedures."
Dr. Marcelo P. Coba (The Wellcome Trust Sanger Institute, Cambridge, UK)

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Selected References for Phosphorylation Site Detector:

Peptide Arrays for Enzyme Profiling
Thiele et al., Methods Mol. Biol. (2009)
Side Chain Specificity of ADP-Ribosylation by a Sirtuin
Fahie et al., FEBS Journal (2009)
High-Density Peptide Microarrays for Reliable Identification of Phosphorylation Sites and Upstream Kinases
Thiele et al., Methods Mol Biol. (2009)
Neurotransmitters Drive Combinatorial Multistate Postsynaptic Density Networks
Coba et al., Science Signaling (2009)

More references under JPT Publications/Literature