Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-(Ser/Thr) PDK1 Docking Motif (18A2) Mouse mAb #9634

Applications Reactivity Sensitivity Isotype
W IP E-P All Endogenous Mouse IgG2a

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/Thr) PDK1 Docking Motif (18A2) Mouse mAb detects phosphorylated serine or threonine that is surrounded by tyrosine or phenylalanine at the -1 and +1 positions and phenylalanine at the -4 position. It also recognizes peptides containing lysine instead of phenylalanine at the -4 position. This antibody does not cross-react with the nonphosphorylated PDK1 docking motifs or with other phosphorylated motifs. This antibody detects endogenous levels of phosphorylated proteins containing PDK1 docking motif, including phospho-Akt. (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 peptides containing the PDK1 docking motif.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or calyculin A-treated (0.1 µM for 30 minutes prior to lysis), using Phospho-(Ser/Thr) PDK1 Docking Motif (18A2) Mouse mAb.

IP

IP

Immunoprecipitation of extracts from NIH/3T3 cells, untreated or treated with 100 ng/ml PDGF for 20 minutes prior to lysis, using Phospho-(Ser/Thr) Docking Motif (18A2) Mouse mAb and Akt Antibody #9272. Western blots were performed using Phospho-(Ser/Thr) Docking Motif (18A2) Mouse mAb (upper) or Akt Antibody #9272 (lower).

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser/Thr) PDK1 Docking Motif (18A2) Mouse mAb ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* and S* denote phosphorylated threonine and serine.)


Background

A hallmark of signal transduction pathways is the reversible phosphorylation of serine and threonine residues within specific sequences, or motifs, in target proteins. Specific signaling motifs include not only sequences that are recognized by protein kinases (1), but also those that are recognized by phosphorylation-dependent binding proteins such as 14-3-3 (2). These modular phosphoprotein interacting domains are critical elements in modulating, directing and amplifying intracellular communications. CST has pioneered the development of phospho-motif specific antibodies, which are invaluable tools for probing the complexity of phospho-regulatory pathways.

Many critical protein kinases can be regulated by phosphorylation at a specific serine or threonine in a hydrophobic motif (3). For example, Akt, a kinase that regulates cell survival, is activated by phosphorylation at Ser473, a site preceded by Phe at -4 and -1 and followed by Tyr at +1 (4). RSK2, p70 S6 kinase and certain PKC isoforms also contain a similar consensus phosphorylation motif. Phosphorylation of these motifs is required for binding to 3-phosphoinositide-dependent kinase 1 (PDK1) (5-7). Phospho-(Ser/Thr) PDK1 Docking Motif (18A2) Monoclonal Antibody is a powerful tool for the characterization of phosphorylated PDK1 docking motifs and the identification of new proteins with PDK1 docking motifs.

  1. Pinna, L.A. and Ruzzene, M. (1996) Biochim Biophys Acta 1314, 191-225.
  2. Yaffe, M.B. and Elia, A.E. (2001) Curr Opin Cell Biol 13, 131-8.
  3. Vanhaesebroeck, B. and Alessi, D.R. (2000) Biochem J 346 Pt 3, 561-76.
  4. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
  5. Frödin, M. et al. (2000) EMBO J 19, 2924-34.
  6. Balendran, A. et al. (1999) J Biol Chem 274, 37400-6.
  7. Balendran, A. et al. (2000) J Biol Chem 275, 20806-13.

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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