Product Pathways - Cell Cycle / Checkpoint
Phospho-CDK2 (Thr160) Antibody #2561
PhosphoSitePlus® protein, site, and accession data: CDK2
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP F E-P | H M R | Endogenous | 33 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
F=Flow Cytometry
E-P=ELISA (Peptide)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Phospho-CDK2 (Thr160) Antibody detects endogenous levels of CDK2 only when phosphorylated at threonine 160. The antibody weakly cross-reacts with cdc2 phosphorylated at Thr161. It does not cross-react with other phosphorylated cyclin dependent kinases.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr160 of human CDK2. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from HeLa cells synchronized in G0, G1/S or Mitosis, using Phospho-CDK2 (Thr160) Antibody.
Flow Cytometry
Flow cytometric analysis of untreated Jurkat cells, using Phospho-CDK2 (Thr160) Antibody versus propidium iodide (DNA content). The boxed population indicates phospho-CDK2 (Thr160)-positive cells.
Background
Cyclin-dependent kinase 2 (p33CDK2) is an important component of the cell cycle machinery. Like p34cdc2, kinase activity is regulated by phosphorylation state as well as association with a cyclin subunit and a CDK inhibitor. Inhibitory phosphorylation occurs on Thr14 and Tyr15 (1). Inhibition of CDK2-cyclin complexes can also be attributed to association with p27 Kip1 and p21 Waf1/Cip1 (2). Activation of CDK2 complexes requires dephosphorylation of Thr14 and Tyr15 by cdc25 phosphatase and phosphorylation of Thr160 (3), which is mediated by CAK, a complex of CDK7 and cyclin H (4). CDK2/cyclin E kinase activity is important for the G1 to S transition and phosphorylation of the Rb protein. During S-phase, active CDK2/cyclin A complexes predominate and phosphorylate E2F and the active CDK2 complex persists in the nucleus throughout G2 (5).
- Morgan, D.O. (1995) Nature 374, 131-134.
- Poon, R.Y. et al. (1996) J. Biol. Chem. 271, 13283-13291.
- Gu, Y. et al. (1992) EMBO J. 11, 3995-4005.
- Fesquet, D. et al. (1993) EMBO J. 12, 3111-3121.
- Morgan, D.O. (1997) Annu. Rev. Cell Dev. Biol. 13, 261-291.
Application References
- Chiariello, M. et al. (2000) Regulation of cyclin-dependent kinase (CDK) 2 Thr-160 phosphorylation and activity by mitogen-activated protein kinase in late G1 phase. Biochem. J. 349, 869-876. Applications: Western Blot
- Lents, N. H. et al. (2002) Stimulation of the Raf/MEK/ERK cascade is necessary and sufficient for activation and Thr-160 phosphorylation of a nuclear-targeted CDK2. J. Biol. Chem. 277, 47469-47475. Applications: Western Blotting
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Companion Products
- 2552 p27 Kip1 Antibody
- 9111 Phospho-cdc2 (Tyr15) Antibody
- 9114 Phospho-cdc2 (Thr161) Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
