Product Pathways - PI3K / Akt Signaling
Phospho-PDK1 (Ser241) Antibody #3061
PhosphoSitePlus® protein, site, and accession data: PDK1
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP | H M R | Endogenous | 58 to 68 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Phospho-PDK1 (Ser241) Antibody detects PDK1 only when phosphorylated at Ser241.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser241 of human PDK1. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from NIH/3T3 cells (starved for 16 hours) treated with PDGF (50 ng/ml), using Phospho-PDK1 (Ser241) Antibody (upper) or control PDK1 antibody (lower). The phospho-specificity of the antibody was confirmed by treating the membrane with calf intestinal alkaline phosphatase (CIP) after Western transfer.
Western Blotting
Western blot analysis of extracts from 293 cells transiently transfected with DNA constructs expressing Wild-type PDK1 or S241A mutant PDK1, treated with IGF-1, using Phospho-PDK1 (Ser241) Antibody (upper) or control PDK1 Antibody #3062 (lower). (Cell lysates provided by Dr. M. Scheid, Ontario Cancer Institute, University Health Network, Toronto.)
Background
Phosphoinositide-dependent protein kinase 1 (PDK1) plays a central role in many signal transduction pathways (1,2) including the activation of Akt and the PKC isoenzymes p70 S6 kinase and RSK (3). Through its effects on these kinases, PDK1 is involved in the regulation of a wide variety of processes, including cell proliferation, differentiation and apoptosis.
Several serine sites (Ser25, Ser241, Ser393/396 and Ser410) are phosphorylated on PDK1 in unstimulated human embryo kidney 293 cells, as well as IGF-1 stimulated cells (4). Phosphorylation on the activation loop Ser241 by autophosphorylation is necessary for PDK1 activity (4).
- Belham, C. et al. (1999) Curr. Biol. 9, R93-R96.
- Toker, A. and Newton, A.C. (2000) Cell 103, 185-188.
- Williams, M.R. et al. (2000) Curr. Biol. 10, 439-448.
- Casamayor, A. et al. (1999) Biochem J 342 ( Pt 2), 287-92.
Application References
- Tanaka, Y. et al. (2003) J Biol Chem 278, 33753-62. Applications: Western Blotting
- Sawitzky, M. et al. (2012) PLoS One 7, e39711. Applications: Western Blotting
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For Research Use Only. Not For Use In Diagnostic Procedures.