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, IHC-P, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

48

SOURCE:

Rabbit

UniProt ID:

#P24864

Entrez-Gene Id:

898

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:100
Flow Cytometry (Fixed/Permeabilized) 1:50

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-Cyclin E1 (Thr62) Antibody detects endogenous levels of cyclin E only when phosphorylated at threonine 62 (cyclin E1 isoform 2) or threonine 77 (cyclin E1 isoform 1).

Species Reactivity:

Human

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr62 of human cyclin E1. Antibodies are purified by protein protein A and peptide affinity chromatography.

Background

Cyclin E1 and cyclin E2 can associate with and activate CDK2 (1). Upon DNA damage, upregulation/activation of the CDK inhibitors p21 Waf1/Cip1 and p27 Kip1 prevent cyclin E/CDK2 activation, resulting in G1/S arrest. When conditions are favorable for cell cycle progression, cyclin D/CDK4/6 phosphorylates Rb and is thought to reduce the activity of p21 Waf1/Cip1 and p27 Kip1, allowing subsequent activation of cyclin E/CDK2 (1,2). Cyclin E/CDK2 further phosphorylates Rb to allow progression into S-phase, where cyclin E/CDK2 is thought to phosphorylate and activate multiple proteins involved in DNA synthesis (2,3). Turnover of cyclin E is largely controlled by phosphorylation that results in SCFFbw7-mediated ubiquitination and proteasome-dependent degradation (4,5). Cyclin E1 is phosphorylated at multiple sites in vivo including Thr62, Ser88, Ser72, Thr380, and Ser384, and is controlled by at least two kinases, CDK2 and GSK-3 (6,7).

  1. Lauper, N. et al. (1998) Oncogene 17, 2637-43.
  2. Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
  3. Ewen, M.E. (2000) Genes Dev 14, 2265-70.
  4. Won, K.A. and Reed, S.I. (1996) EMBO J 15, 4182-93.
  5. Koepp, D.M. et al. (2001) Science 294, 173-7.
  6. Welcker, M. et al. (2003) Mol Cell 12, 381-92.
  7. Ye, X. et al. (2004) J Biol Chem 279, 50110-9.

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 IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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
#4136

Phospho-Cyclin E1 (Thr62) Antibody

Western Blotting Image 1: Phospho-Cyclin E1 (Thr62) Antibody Expand Image
Western blot analysis of extracts from 293 cells, treated with 10 mM LiCl for the indicated times, using Phospho-Cyclin E1 (Thr62) Antibody (upper) or a cyclin E antibody (lower).
Western Blotting Image 2: Phospho-Cyclin E1 (Thr62) Antibody Expand Image
Western blot analysis of extracts from 293 cells, untreated or hydroxyurea-treated, using Phospho-Cyclin E1 (Thr62) Antibody.
No image available
Immunohistochemistry Image 1: Phospho-Cyclin E1 (Thr62) Antibody Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing nuclear localization, using Phospho-Cyclin E1 (Thr62) Antibody.
Immunohistochemistry Image 2: Phospho-Cyclin E1 (Thr62) Antibody Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-Cyclin E1 (Thr62) Antibody.
Immunohistochemistry Image 3: Phospho-Cyclin E1 (Thr62) Antibody Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-Cyclin E1 (Thr62) Antibody in the presence of control peptide (left) or antigen-specific peptide (right).
Flow Cytometry Image 1: Phospho-Cyclin E1 (Thr62) Antibody Expand Image
Flow cytometric analysis of untreated Jurkat cells, using Phospho-Cyclin E1 (Thr62) Antibody versus propidium iodide (DNA content). The boxed population indicates phospho-Cyclin E (Thr62)-positive cells.