Revision 1

#9631Store at -20C

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:

Rabbit

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
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.

Specificity / Sensitivity

Phospho-(Ser/Thr) Phe Antibody detects phospho-serine or threonine in the context of tyrosine, tryptophan or phenylalanine at the -1 position or phenylalanine at the +1 position. The antibody does not cross-react with the nonphosphorylated form of these motifs, nor does it cross-react with other phospho-serine/threonine-containing proteins and peptides. (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.)

Species Reactivity:

All Species Expected

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phospho-serine/threonine-phenylalanine-containing peptide. Antibodies are purified by protein A and peptide affinity chromatography.

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 surrounded by phenylalanine or tyrosine. For example, Akt, a kinase that regulates cell survival, is activated by phosphorylation at Ser473, a site surrounded by phenylalanine and tyrosine (3). RSK1, p70S6K, and certain PKC isoforms also contain a similar consensus phosphorylation site. Phosphorylation of these sites is required for kinase activity (4,5). The Phospho-(Ser/Thr) Phe Antibody is a powerful tool for discovery of new proteins containing this important regulatory motif.

  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. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
  4. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
  5. Keranen, L.M. et al. (1995) Curr Biol 5, 1394-1403.

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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#9631

Phospho-(Ser/Thr) Phe Antibody

Western Blotting Image 1: Phospho-(Ser/Thr) Phe Antibody Expand Image
Western blot analysis of extracts from A431 cells, untreated or calyculin A-treated, using Phospho-(Ser/Thr) Phe Antibody.
No image available
Product Image 1: Phospho-(Ser/Thr) Phe Antibody Expand Image
Phospho-(Ser/Thr) Phe Antibody ELISAs: Signal to noise ratio of phospho- versus nonphospho-peptides in which the phosphorylation site is adjacent to phenylalanine, tyrosine or tryptophan. (T* and S* denote phosphorylated threonine and serine.)