Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Cell Cycle / Checkpoint

Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb (PE Conjugate) #8497

Applications Reactivity Sensitivity Isotype
F H (Mk) Endogenous Rabbit IgG

Applications Key:  F=Flow Cytometry
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

* Product-specific protocol.

Specificity / Sensitivity

Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb (PE Conjugate) detects endogenous levels of cyclin D1 only when phosphorylated at Thr286. This antibody does not cross-react with other cyclin D family members.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr286 of cyclin D1 protein.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HT-1080 cells, MG132 and λ phosphatase-treated (red), untreated (blue), or MG132-treated only (green), using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb (PE Conjugate).

Description

This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometry analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb #3300.

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 affect progression through the restriction point and Rb-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 protein expression and phosphorylation-dependent degradation (4).

  1. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
  2. Sherr, C.J. (1996) Science 274, 1672-1677.
  3. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
  4. Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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