Product Pathways - Motif Antibodies

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Phospho-PKA Substrate (RRXS/T) (100G7E) Rabbit mAb #9624

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Applications	Reactivity	Sensitivity	Isotype
W IP E-P	All	Endogenous	Rabbit IgG

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

9624:: ELISA Peptide, Immunoprecipitation, Western Blotting

Specificity / Sensitivity

Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb detects peptides and proteins containing a phospho-Ser/Thr residue with arginine at the -3 and -2 positions. It is a useful tool in identifying new substrates of PKA. The antibody recognizes other -3 arginine-bearing phospho-Ser/Thr peptides, such as substrate motifs for Akt and PKC, to a lesser extent. It does not recognize the nonphosphorylated substrate motif peptides. (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

Monoclonal antibody is produced by immunizing animals with synthetic phospho-PKA substrate peptides .

Western Blotting

Western blot analysis of extracts from serum-starved A431 cells, phosphorylated in vitro with PKA kinase or treated in culture with forskolin/IBMX, using Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb. Lysis buffer: 1.0% Triton X-100 (lanes 1 and 2), 2.0% SDS (lanes 3 and 4).

ELISA-Peptide

Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* and S* denote phosphorylated threonine and serine.)

Background

An important class of kinases, refered to as Arg-directed kinases or AGC-family kinases, includes cAMP-dependent protein kinase (PKA), cGMP-dependent protein kinase (PKG), protein kinase C, Akt and RSK. These kinases share a substrate specificity characterized by Arg at position -3 relative to the phosphorylated Ser or Thr (1,2). Phospho-PKA substrate-specific antibodies from Cell Signaling Technology are powerful tools for investigating the regulation of phosphorylation by PKA and other Arg-directed kinases, as well as for high throughput kinase drug discovery.

Application References

Bradshaw, N.J. et al. (2011) J Neurosci 31, 9043-54. Applications: Western Blotting
Park, J.H. et al. (2011) J Biol Chem 286, 23132-41. Applications: Western Blotting
Biton, S. and Ashkenazi, A. (2011) Cell 145, 92-103. Applications: IP
Li, X. et al. (2006) Biochem J 394, 427-35. Applications: Western Blotting
Moir, R.D. et al. (2006) Proc Natl Acad Sci U S A 103, 15044-9. Applications: Western Blotting

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Companion Products

Rabbit Monoclonals Produced Using Technology from Epitomics, Inc. Under U.S. patent No. 5,675,063.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.