Product Pathways - Motif Antibodies
Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb #9614
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| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| W IP IHC-P E-P | All | Endogenous | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
E-P=ELISA (Peptide)
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.
Protocols
Specificity / Sensitivity
Phospho-(Ser/Thr) Akt Substrate Motif (RXXS*/T*)(110B7) RmAb recognizes peptides and proteins containing phospho-serine/threonine preceded by arginine at the -3 position. There is some preference observed for peptides that contain phospho-serine/threonine preceded by arginine at both positions -5 and -3. (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-Akt substrate peptides .
Western Blotting
Western blot analysis of extracts from serum-starved NIH/3T3 cells, phosphorylated in vitro with Akt kinase, or treated in culture with PDGF or FBS/Calyculin A, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
IP
Immunoprecipitation of extracts from serum-starved NIH/3T3 cells, untreated or treated with FBS/Calyculin A, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb, followed by Western blot analysis using the same antibody.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma control (left) or lambda phosphatase-treated (right), using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded lung carcinoma, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded renal cell carcinoma, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
ELISA-Peptide
Phospho-Akt Substrate (RXXS*/T*) (110B7E) 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, referred 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). Akt plays a central role in mediating critical cellular responses including cell growth and survival, angiogenesis, and transcriptional regulation (3-5). While a number of Akt substrates are known (such as GSK-3, Bad, and caspase-9) many important substrates await discovery. Akt phosphorylates substrates only at Ser/Thr in a conserved motif characterized by Arg at positions -5 and -3 (6). Phospho-Akt substrate-specific antibodies from Cell Signaling Technology are powerful tools for investigating the regulation of phosphorylation by Akt and other Arg-directed kinases, as well as for high throughput kinase drug discovery.
- Montminy, M. (1997) Annu Rev Biochem 66, 807-22.
- Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
- Marte, B.M. and Downward, J. (1997) Trends Biochem Sci 22, 355-8.
- Jiang, B.H. et al. (2000) Proc Natl Acad Sci USA 97, 1749-53.
- Scheid, M.P. and Woodgett, J.R. (2000) Curr Biol 10, R191-4.
- Alessi, D.R. et al. (1996) FEBS Lett 399, 333-8.
Application References
- Fonseca, B.D. et al. (2011) J Biol Chem 286, 27111-22. Applications: Western Blotting
- Koumanov, F. et al. (2011) J Biol Chem 286, 16574-82. Applications: Western Blotting
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
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- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 6211 SignalSilence® Akt siRNA I
- 6510 SignalSilence® Akt siRNA II
- 6909 SignalSilence® Akt1 siRNA I (Mouse Specific)
- 6396 SignalSilence® Akt2 siRNA
- 6407 SignalSilence® Akt2 siRNA I (Mouse Specific)
- 6408 SignalSilence® Akt2 siRNA II (Mouse Specific)
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.
