Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-(Ser) PKC Substrate Antibody #2261

Applications Reactivity Source
W IP E-P All Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected

Specificity / Sensitivity

Phospho-(Ser) PKC Substrate Antibody detects endogenous levels of many cellular proteins only when phosphorylated at serine residues surrounded by Arg or Lys at the -2 and +2 positions and a hydrophobic residue at the +1 position. The antibody does not cross-react with nonphosphorylated serine residues, with phospho-threonine in the same motif, or with phospho-serine in other motifs. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic phospho-PKC substrate peptides (KLH-coupled). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or treated with PMA (50 ng/ml) and ionomycin (1 µM for 20 minutes prior to lysis) and subjected to 2-D electrophoresis, using Phospho-(Ser) PKC Substrate Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from WEHI 231 cells and Jurkat cells, untreated or TPA-treated, using Phospho-(Ser) PKC Substrate Antibody.

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser) PKC Substrate Antibody ELISA assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (S* denotes phosphorylated serine.)


Background

Although protein kinase C (PKC) family members are involved in a number of signal transduction processes including secretion, gene expression, proliferation and muscle contraction (1,2), many of their substrates remain unidentified. Isozymes of PKC are subdivided into conventional PKCs (cPKC), novel PKCs (nPKC) and atypical PKCs (aPKC). PKCalpha, beta I, beta II and gamma isoforms belong to cPKC (1). When activated, cPKC isozymes phosphorylate substrates containing serine or threonine, with arginine or lysine at the -3, -2 and +2 positions, and hydrophobic amino acids at position +1 (1-3). To identify potential PKC substrates, CST has developed antibodies that recognize this motif. As shown by ELISA, Phospho-(Ser) PKC Substrate Antibody is specific to cPKC substrates containing phospho-serine.

  1. Nishikawa, K. et al. (1997) J Biol Chem 272, 952-60.
  2. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
  3. Obata, T. et al. (2000) J Biol Chem 275, 36108-15.

Application References

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Companion Products

License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.'s 7,198,896 & 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 commerical licensing terms please contact CST Business Development at cbunker@cellsignal.com.

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