Product Pathways - Cell Cycle / Checkpoint
Phospho-Cyclin D1 (Thr286) Antibody #2921
| Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|
| W | H R (M) | 36 | Rabbit |
Applications Key:
W=Western Blotting
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-Cyclin D1 (Thr286) Antibody detects endogenous levels of cyclin D1 only when phosporylated at threonine 286. The antibody does not cross-react with other cyclin D family members at physiological levels.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr286 of human cyclin D1. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from SK-N-MC and C6 cells, untreated or EGF-treated, using Phospho-Cyclin D1 (Thr286) Antibody (upper) or Cyclin D1 mouse mAb #2926 (lower).
Western Blotting
Western blot analysis of extracts from SK-N-MC cells, untreated or calf intestinal phosphatase (CIP)-treated, using Phospho-Cyclin D1 (Thr286) Antibody (upper) or Cyclin D1 Mouse mAb #2926 (lower).
Background
Activity of the cyclin-dependent kinases CDK4 and CDK6 is regulated by T-loop phosphorylation, by the abundance of their cyclin partners (the D-type cyclins), and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of cyclin D/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the release and degradation of p27 concomitant with a rise in cyclin D levels to effect progression through the restriction point and pRb-dependent entry into S-phase (2). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). Levels of cyclin D protein drop upon withdrawal of growth factors through downregulation of its protein expression and through phosphorylation-dependent degradation (4).
Ubiquitination of cyclin D1 is enhanced by phosphorylation at Thr286 by glycogen synthase kinase 3beta (GSK-3beta) (4).
- Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
- Sherr, C.J. (1996) Science 274, 1672-1677.
- Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
- Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.
Application References
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