antibody

Cyclin E, phosphorylated (Thr62)

C8455-23

100 ul

polyclonal

domestic rabbit

nonconjugated

phosphorylated

Cyclin E, phosphorylated (Thr62)

rabbit

Pab

Phosphorylated

Antibodies

100 ul

IgG

Cyclin E1 and cyclin E2 can associate with and activate cdk2 (1). Upon DNA damage, upregulation/ activation of the cdk inhibitors p21Waf1/Cip1 and p27Kip1 prevents cyclin E/cdk2 activation, resulting in G1/S arrest. However, when conditions are favorable for cell cycle progression, cyclin D/cdk4/6 phosphorylates Rb and is thought to reduce the activity of p21Waf1/Cip1 and p27Kip1, allowing subsequent activation of cyclin E/cdk2 (1,2). Cyclin E/cdk2 then 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 contolled by phosphorylation resulting in SCFFbw7-mediated ubiquitination and proteasomedependent 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 GSK3 (6,7). Applications: Suitable for use in Flow Cytometry, Western Blot, Immunoprecipitation, Immunohistochemistry. Other applications not tested. Recommended Dilution: Flow Cytometry: 1:25 Western Blot: 1:1000 Immunohistochemistry (Paraffin) 1:100 Citrate/TBST Immunoprecipitation: 1:100 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot. Store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Hu

Synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr62 of human cyclin E.

Detects endogenous levels of cyclin E only when phosphorylated at threonine 62 (cyclin E1 isoform 2) or threonine 77 (cyclin E1 isoform 1). Species Crossreactivity: Human

Purified by Protein A and peptide affinity chromatography

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol.

100ul (10 Western mini-blots)

Product Type: Pab Host: rabbit Source: human Concentration: 100ul (10 Western mini-blots) Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM NaCl, 100ug/ml BSA, 50% glycerol. Purity: Protein A and peptide affinity chromatography Immunogen: Synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr62 of human cyclin E. Specificity: Detects endogenous levels of cyclin E only when phosphorylated at threonine 62 (cyclin E1 isoform 2) or threonine 77 (cyclin E1 isoform 1). Species Crossreactivity: Human Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

FC IHC IP WB

-20°C

(1) Lauper, N. et al. (1998) Oncogene 17, 2637–2643. (2) Lundberg, A.S. and Weinberg, R.A. (1998) Mol. Cell. Biol. 18, 753–761. (3) Ewen, M.E. (2000) Genes Dev. 14, 2265–2270. (4) Won, K.A. and Reed, S.I. (1996) EMBO J. 15, 4182–4193. (5) Koepp, D.M. et al. (2001) Science 294, 173–177. (6) Welcker, M. et al. (2003) Mol. Cell 12, 381–392. (7) Ye, X. et al. (2004) J. Biol. Chem. 279, 50110–50119.

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