Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, E-P

REACTIVITY:

All

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Mouse IgG2a

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Peptide ELISA (DELFIA) 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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.

Species Reactivity:

All Species Expected

Source / Purification

Monoclonal antibody is produced by immunizing animals with peptides containing the PDK1 docking motif.

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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation E-P: Peptide ELISA (DELFIA)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专