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, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

57

SOURCE:

Rabbit

UniProt ID:

#P49918

Entrez-Gene Id:

1028

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:100

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-p57 Kip2 (Thr310) Antibody detects endogenous levels of p57 Kip2 only when phosphorylated at threonine 310. This antibody may cross-react with p27 Kip1 when phosphorylated at Thr187.

Species Reactivity:

Human

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr310 of human p57 Kip2. Antibodies are purified by protein A and peptide affinity chromatography.

Background

p27 Kip1 is a member of the Cip/Kip family of cyclin-dependent kinase inhibitors. Like its relatives, p57 Kip2 and p21 Waf1/Cip1, the ability to enforce the G1 restriction point is derived from its inhibitory binding to CDK2/cyclin E and other CDK/cyclin complexes. Expression levels of p27 are upregulated in quiescent cells and in cells treated with cAMP or other negative cell cycle regulators. Downregulation of p27 can be induced by treatment with interleukin-2 or other mitogens; this involves phosphorylation of p27 and its degradation by the ubiquitin-proteasome pathway (1-4).
Levels of p57 Kip2 are controlled by ubiquitination/degradation via the Skp1/Cul1/F-box-type E3 ubiquitin ligase complex SCF-Skp2, and this effect is dependent on Thr310 (5). A similar threonine phosphorylation site in p27 Kip1, Thr187, has also been shown to regulate protein stability (6).

  1. Lloyd, R.V. et al. (1999) Am J Pathol 154, 313-23.
  2. Polyak, K. et al. (1994) Genes Dev 8, 9-22.
  3. Kato, J.Y. et al. (1994) Cell 79, 487-96.
  4. Vlach, J. et al. (1997) EMBO J 16, 5334-44.
  5. Kwon, T. et al. (2000) J Biol Chem 275, 423-8.
  6. Heeneman, S. et al. (2000) J Biol Chem 275, 15926-32.

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 IF-IC: Immunofluorescence (Immunocytochemistry)

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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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
#2558

Phospho-p57 Kip2 (Thr310) Antibody

Western Blotting Image 1: Phospho-p57 Kip2 (Thr310) Antibody Expand Image
Western blot analysis of extracts from untreated and dexamethasone-treated (50 nM, 16 h) HeLa cells, using Phospho-p57 Kip2 (Thr310) Antibody (upper) and p57 Kip2 Antibody #2557 (lower). Lysates were treated with lambda phosphatase NEB#P0753 (10,000 U/ml for 30 mins) to confirm the phospho-specificity of the phospho-antibody.
Immunofluorescence Image 1: Phospho-p57 Kip2 (Thr310) Antibody Expand Image
Confocal immunofluorescent analysis of HeLa cells, dexamethasone-treated (left) or phosphatase-treated (right), labeled with Phospho-p57 Kip2 (Thr310) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).