Phospho-(Ser/Thr) Akt Substrate Antibody #9611
Filter:
- WB
- IP
- IHC
Supporting Data
REACTIVITY | All |
SENSITIVITY | Endogenous |
MW (kDa) | |
SOURCE | Rabbit |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IHC-Immunohistochemistry
Species Cross-Reactivity Key:
- All-All Species Expected
Product Information
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Immunohistochemistry (Paraffin) | 1:500 |
Peptide ELISA (DELFIA) | 1:500 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
Protocol
Specificity / Sensitivity
Phospho-(Ser/Thr) Akt Substrate Antibody preferentially recognizes peptides and proteins containing phospho-Ser/Thr preceded by Lys/Arg at positions -5 and -3, in a manner largely independent of other surrounding amino acids. Some cross-reactivity is observed for peptides that contain phospho-Ser/Thr preceded by Arg/Lys at positions -3 and -2. No cross-reactivity is observed with the corresponding nonphosphorylated sequences or with other phospho-Ser/Thr-containing motifs. By ELISA, the antibody recognizes a wide range of phosphorylated Akt substrate peptides, and, by 2-D gel Western blot analysis, it recognizes a large number of proteins presumed to be Akt substrates.
Species Reactivity:
All Species Expected
Source / Purification
Polyclonal antibodies are produced by immunizing animals with phospho-Akt substrate peptides . Antibodies are purified by protein A and peptide affinity chromatography.
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.
限制使用
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For Research Use Only. Not For Use In Diagnostic Procedures.
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