Product Pathways - Cell Cycle / Checkpoint
p27 Kip1 (D69C12) XP® Rabbit mAb #3686
PhosphoSitePlus® protein, site, and accession data: p27Kip1
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IF-IC F | H R Mk | Endogenous | 27 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
R=Rat
Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
p27 Kip1 (D69C12) XP® Rabbit mAb detects endogenous levels of total p27 Kip1 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human p27 Kip1.
Western Blotting
Western blot analysis of extracts from various cell types using p27 Kip1 (D69C12) XP® Rabbit mAb.
IP
Immunoprecipitation of p27 Kip1 from 293 cells using p27 Kip1 (D69C12) XP® Rabbit mAb. Western analysis was performed using the same antibody. Lane 1 is 5% input.
Flow Cytometry
Flow cytometric analysis of Jurkat cells using p27 Kip1 (D69C12) XP® Rabbit mAb versus Propidium Iodide (PI)/RNase Staining Solution #4087. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
IF-IC
Confocal immunofluorescent analysis of MCF-7 cells using p27 Kip1 (D69C12) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Background
p27 Kip1 is a member of the Cip/Kip family of cyclin-dependent kinase inhibitors. Like its relatives, p57 Kip2 and p21 Waf1/Cip1, the ability to enforce the G1 restriction point is derived from its inhibitory binding to CDK2/cyclin E and other CDK/cyclin complexes. Expression levels of p27 are upregulated in quiescent cells and in cells treated with cAMP or other negative cell cycle regulators. Downregulation of p27 can be induced by treatment with interleukin-2 or other mitogens; this involves phosphorylation of p27 and its degradation by the ubiquitin-proteasome pathway (1-4).
- Lloyd, R.V. et al. (1999) Am. J. Pathol. 154, 313-323.
- Polyak, K. et al. (1994) Genes Dev. 8, 9-22.
- Kato, J.Y. et al. (1994) Cell 79, 487-496.
- Vlach, J. et al. (1997) EMBO J. 16, 5334-5344.
Application References
- Stein, C. et al. (2012) Nucleic Acids Res , . Applications: Western Blotting
- Poindessous, V. et al. (2011) Clin Cancer Res 17, 6522-30. Applications: Western Blotting
- Yang, S. et al. (2011) PLoS One 6, e26343. Applications: Western Blotting
- Negrotto, S. et al. (2011) Cancer Res 71, 1431-41. Applications: Western Blotting
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Companion Products
- 2552 p27 Kip1 Antibody
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- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked 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.
