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9615
Phospho-(Ser) Kinase Substrate Antibody Sampler Kit

Phospho-(Ser) Kinase Substrate Antibody Sampler Kit #9615

Western Blotting Image 1

Western blot analysis of extracts from serum-starved A431 cells, untreated or treated in culture with EGF, and HeLa cells, untreated, hydroxyurea-treated (G1/S) or Nocodazole-treated (G2/M), using Phospho-MAPK/CDK Substrate (PXS*P or S*PXR/K) (34B2) Rabbit mAb.

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Western Blotting Image 2

Western blot analysis of extracts from A431 cells, untreated or calyculin A treated, using Phospho-(Ser) Arg-X-Tyr/Phe-X-pSer Motif Antibody.

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Western Blotting Image 3

Western blot analysis of extracts from A431 cells, untreated or calyculin A-treated, using Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb (left) or Phospho-(Ser) 14-3-3 Binding Motif Antibody #9601 (right).

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Western Blotting Image 4

Western blot analysis of extracts from COS-7 cells, synchronized in G1/S, G2/M, and G0 phase of the cell cycle, using

Phospho-CDK Substrate Motif [(K/H)pSP] MultiMab™ Rabbit mAb mix.

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Western Blotting Image 5

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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IHC-P (paraffin) Image 6

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, control (left) or lambda phosphatase-treated (right), using Phospho-MAPK/CDK Substrates (PXS*P or S*PXR/K) (34B2) Rabbit mAb.

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IHC-P (paraffin) Image 7

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-MAPK/CDK Substrate (PXS*P or S*PXR/K) (34B2) Rabbit mAb.

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IHC-P (paraffin) Image 8

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-MAPK/CDK Substrate (PXS*P or S*PXR/K) (34B2) Rabbit mAb.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-MAPK/CDK Substrates (PXS*P or S*PXR/K) (34B2) Rabbit mAb 2325 20 µl
  • WB
  • IP
  • IHC
H All Rabbit IgG
Phospho-(Ser) Arg-X-Tyr/Phe-X-pSer Motif Antibody 2981 20 µl
  • WB
  • IP
H M All Rabbit 
Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb 9606 20 µl
  • WB
  • IP
H All Mouse IgG1
Phospho-CDK Substrate Motif [(K/H)pSP] MultiMab™ Rabbit mAb mix 9477 20 µl
  • WB
  • IP
H All Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

The Phospho-(Ser) Kinase Substrate Antibody Sampler Kit provides a fast and economical means of evaluating several phospho-Ser kinase substrates. The kit contains enough primary and secondary antibody to perform two Western blot experiments.

Each antibody detects endogenous levels of phospho-(Ser) proteins of specific kinase substrate groups. For more detailed information please see datasheet for this product.

Polyclonal antibodies are produced by immunizing animals with phosphopeptides containing the kinase substrate motif and purified by protein A and peptide affinity chromatography.

Monoclonal antibody is produced by immunizing animals with synthetic phospho-MAPK/CDK substrate and phospho-CDKs substrate peptides.

Mouse monoclonal antibody is produced by immunizing animals with phospho-(Ser) 14-3-3 binding motif peptides .

Phospho-(Ser) kinases and phosphatases play critical roles in a wide range of biological processes. Each phospho-(Ser) kinase phosphorylates serine within a specific motif. The MAPK and CDK families of serine protein kinases phosphorylate serine followed by proline residue (1-3). The consensus amino acid sequence for CDK substrate is (K/R)(S*)PX(K/R), where X denotes any one of the 20 amino acids and S* is the phosphorylation site (4-6). MAPK phosphorylates substrates with the concensus sequence PX(S*)P. The 14-3-3 proteins are a highly conserved family of proteins involved in the regulation of cell survival, apoptosis, proliferation and checkpoint control (7-11). Binding of 14-3-3 is mediated through phospho-serine-containing proteins (12). Two different phospho-serine containing motifs are found using a degenerate phospho-serine-oriented peptide library technique, RSXS*XP and RXY/FXS*XP (12). Motif 1 (Arg/Lys and Ser at positions -3 and -2, phospho-Ser at position 0, and Pro at position +2) is found in critical regulatory proteins including Bad, cdc25C, FoxO3A, PKC and c-Raf (11, 13). Motif 2 (RXY/FXS*XP) is found in critical regulatory proteins including cdc25A, cdc25B, PKCγ, IRS-1 and BCR (12). Although Phospho-(Ser) Arg-X-Tyr/Phe-X-pSer Motif Antibody binds 14-3-3 binding motif 2 with no requirement for proline in the +2 position, it provides a powerful tool for the discovery and characterization of potential 14-3-3 binding motif 2-containing proteins or other proteins with the RXY/FXS* motif. Antibodies specific to particular kinase substrates are invaluable reagents in determining kinase activity and identifying potential new kinase substrates. CST has developed antibodies that recognize phosphorylated serine within the context of a protein motif that is phosphorylated by MAPK/CDK, CDKs or 14-3-3. As shown by DELFIA or ELISA, each phospho-(Ser) kinase substrate antibody in this sampler kit is specific to its kinase substrate motif.

  1. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
  2. Karin, M. (1994) Curr Opin Cell Biol 6, 415-24.
  3. Lewis, T. S. et al. (1998) Adv. Cancer Res. 74, 49-139.
  4. Songyang, Z. et al. (1996) Mol Cell Biol 16, 6486-93.
  5. Songyang, Z. (1999) Prog Biophys Mol Biol 71, 359-72.
  6. Holmes, J.K. and Solomon, M.J. (1996) J Biol Chem 271, 25240-6.
  7. Aitken, A. (1995) Trends Biochem Sci 20, 95-7.
  8. Zha, J. et al. (1996) Cell 87, 619-28.
  9. Piwnica-Worms, H. (1999) Nature 401, 535, 537.
  10. Tzivion, G. et al. (1998) Nature 394, 88-92.
  11. Xing, H. et al. (2000) EMBO J 19, 349-58.
  12. Muslin, A.J. et al. (1996) Cell 84, 889-97.
  13. Yaffe, M.B. et al. (1997) Cell 91, 961-71.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

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