Description
Product Usage Information
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.
Protocol
Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin), Flow Cytometry (Fixed/Permeabilized)
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 including Thr62, Ser88, Ser72, Thr380, and Ser384, and is controlled by at least two kinases, CDK2 and GSK-3 (6,7).
Alternate Names
CCNE; CCNE1; cyclin E1; cyclin Es; cyclin Et; G1/S-specific cyclin-E1; pCCNE1
