Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-(Ser) PKC Substrate Antibody #2261

Applications Reactivity Sensitivity Source
W IP E-P All Endogenous Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

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 animals with synthetic phospho-PKC substrate peptides . 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). PKCα, βI, βII, and γ isoforms belong to the cPKC group (1). When activated, cPKC isozymes phosphorylate substrates containing Ser or Thr, with Arg or Lys at the -3, -2, and +2 positions, and a hydrophobic amino acid at position +1 (1-3). To identify potential PKC substrates, Cell Signaling Technology has developed antibodies that recognize this motif. Phospho-(Ser) PKC Substrate (D13C5/D47H10/D50E3) Rabbit mAb is specific to cPKC substrates containing phospho-Ser.

  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 commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

For Research Use Only. Not For Use In Diagnostic Procedures.

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