Product Pathways - Cell Cycle / Checkpoint
Phospho-cdc2 (Thr161) Antibody #9114
| Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|
| W IP IF-IC E-P | H M R | 34 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
E-P=ELISA (Peptide)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-cdc2 (Thr161) Antibody detects endogenous levels of cdc2 only when phosphorylated at threonine 161. The antibody cross-reacts with endogenous CDK2 phosphorylated at threonine 160.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr161 of human cdc2. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of cdc2 kinase treated with lambda protein phosphatase (lambda-PPase) for the indicated times, using Phospho-cdc2 (Thr161) Antibody.
Western Blotting
Western blot analysis of extracts from HeLa cells synchronized in G0, G1/S, or M phase, using Phospho-cdc2 (Thr161) Antibody
IF-IC
Dual immunofluorescent analysis of proliferating HeLa cells, using cdc2 (POH1) mAb #9116 (left) or Phospho-cdc2 (Thr161) Antibody (right).
Background
The entry of eukaryotic cells into mitosis is regulated by cdc2 kinase activation, a process controlled at several steps including cyclin binding and phosphorylation of cdc2 at Thr161 (1). However, the critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Tyr15 and Thr14 (2). Phosphorylation at Thr14 and Tyr15 resulting in inhibition of cdc2 can be carried out by Wee1 and Myt1 protein kinases (3,4). The cdc25 phosphatase may be responsible for removal of phosphates at Thr14 and Tyr15 and subsequent activation of cdc2 (1,5).
cdc2 activation and association with cyclin A require phosphorylation at Thr161 by the CDK-activating kinase CAK, a complex of CDK7 and cyclin H (7,8).
- Atherton-Fessler, S. et al. (1994) Mol. Biol. Cell. 5, 989-1001.
- Norbury, C. et al. (1991) EMBO. J. 10, 3321-3329.
- McGowan, C.H. and Russell, P. (1993) EMBO J. 12, 75-85.
- Wells, N.J. et al. (1999) J. Cell. Sci. 112, 3361-3371.
- Hunter, T. (1995) Cell 80, 225-236.
- Fesquet, D. et al. (1993) EMBO J. 12, 3111-3121.
- Ducommun, B. et al. (1991) EMBO J. 10, 3311-3319.
Application References
- De Smedt, V. et al. (2002) Thr-161 phosphorylation of monomeric Cdc2. Regulation by protein phosphatase 2C in Xenopus oocytes. J. Biol. Chem. 277, 28592-28600. This article references the use of Phospho-cdc2 (Thr161) Antibody in the following applications: Western Blotting
- Baus, F. et al. (2003) Permanent cell cycle exit in G2 phase after DNA damage in normal human fibroblasts. EMBO J. 22, 3992-4002. This article references the use of Phospho-cdc2 (Thr161) Antibody in the following applications: Western Blotting
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Companion Products
- 9110 PhosphoPlus® cdc2 (Tyr15) Antibody Kit
- 9111 Phospho-cdc2 (Tyr15) Antibody
- 9112 cdc2 Antibody
- 9116 cdc2 (POH1) Mouse mAb
- 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