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You are here:Home » Antibodies » Abs to Protein Kinase Inhibitors » Anti -p27kip1, phosphophorylated (Thr187)

Anti -p27kip1, phosphophorylated (Thr187)

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Specifications

Clone Host Grade Applications
Polyclonal Rabbit Affinity Purified E B
The cyclin-dependent kinase inhibitor p27KIP1 is a potent inhibitor of both Cyclin E-Cdk2 and Cyclin A-Cdk2, two cdk complexes involved in the regulation of the G1/S transition (reviewed in ref 2). p27KIP1 has been found to be expressed at high levels in quiescent cells, playing an important role in maintaining cells in G0 through the inhibition of Cdks. Several antimitogenic
stimuli, for example cell contact and lack of cell adhesion, upregulate p27KIP1 levels. When cells re-enter the cell cycle the abundance of p27KIP1 protein, but not of the mRNA, decreases because of an acceleration of p27KIP1 ubiquitinmediated degradation.(3) Cdk-mediated phosphorylation of p27 on threonine 187 is essential for p27 ubiquitination.(1). Furthermore, Skp2, a human F-box protein essential for entry into S phase, recognizes in vivo p27 only when
phosphorylated on T187 and is a rate-limiting component of the machinery that ubiquitinates p27.(4) Overexpression of p27KIP1 arrests the cell division cycle in G1. Lack of p27KIP1 in mice induces adenomas of the pituitary pars intermedia with 100% penetrance.(5-7) In addition, all the fully developed organs show generalized hyperplasia. This phenotype underscores the importance of p27KIP1 in regulating cell proliferation. In human tumors, lack of p27KIP1 is a powerful independent indicator of a poor clinical prognosis.(8,9)
Catalog #P1001-11
ApplicationsSuitable for use in ELISA, Western Blot and Immunoprecipitation. Other applications not tested.
Recommended DilutionELISA: 0.1-1.0ug/ml. Milk-derived blocking solutions reportedly contain phosphoproteins that may inhibit phosphoamino acid antibody binding and therefore should be avoided. A 3% BSA
solution may also be used.
Immunoprecipitation(1): 15ug/IP reaction
Western Blotting(1): 5-10ug/ml
Optimal dilution determined by the researcher.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Clone TypePolyclonal
HostRabbit
SourceHuman
Concentration0.25 mg/ml
FormSupplied as a liquid in PBS, pH 7.4, containing 0.05% sodium azide.
PurityPurified by affinity chromatography.
ImmunogenThreonine187-phosphorylated p27 peptide derived from the C-terminus of the human p27 protein(1). This sequence differs by
a single amino acid (not at the phosphorylated threonine) in the mouse, rat, and hamster p27 proteins.
SpecificitySpecific for the Thr-187 phosphorylated form of the cyclin-dependent kinase inhibitor p27KIP1. The antibody does not react with unphosphorylated p27 or with related CDK inhibitor proteins. This antibody can be used to detect endogenous phosphorylated-p27 by Western blotting if lysates are first immunoprecipitated with “plain” p27 antibody eg. Rb x p27, cat no. 71-9600 or Ms x p27, cat no.33-2800). Characterization of this antibody has been carried out primarily with purified recombinant p27 incubated in kinase reactions.(1)
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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