Anti-p27 KIP 1 (phospho S10) antibody [EP233(2)Y] (ab62364)
- Product nameAnti-p27 KIP 1 (phospho S10) antibody [EP233(2)Y]See all p27 KIP 1 primary antibodies ...
- DescriptionRabbit monoclonal [EP233(2)Y] to p27 KIP 1 (phospho S10)
- Specificityab62364 detects p27 KIP 1 phosphorylated at Serine 10.
- Tested applicationsWB, IP, IHC-P more details
- Species reactivityReacts with: Mouse, Rat, Human, Monkey
A synthetic phosphopeptide corresponding to residues surrounding Serine 10 of human p27 KIP 1.
- Positive control
- MCF-7 cell lysates, human ovarian carcinoma tissue and human tonsil tissue.
- General notesProduced under U.S. Patent No. 5,675,063.
- Storage instructionsStore at -20°C. Stable for 12 months at -20°C
- Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
- PurityTissue culture supernatant
- Clonality Monoclonal
- Clone numberEP233(2)Y
- Research Areas
Our Abpromise guarantee covers the use of ab62364 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||WB: 1/10000 - 1/20000. Detects a band of approximately 27 kDa (predicted molecular weight: 22 kDa).|
|IHC-P||IHC-P: 1/200 - 1/1500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
- FunctionImportant regulator of cell cycle progression. Involved in G1 arrest. Potent inhibitor of cyclin E- and cyclin A-CDK2 complexes. Forms a complex with cyclin type D-CDK4 complexes and is involved in the assembly, stability, and modulation of CCND1-CDK4 complex activation. Acts either as an inhibitor or an activator of cyclin type D-CDK4 complexes depending on its phosphorylation state and/or stoichometry.
- Tissue specificityExpressed in all tissues tested. Highest levels in skeletal muscle, lowest in liver and kidney.
- Involvement in diseaseDefects in CDKN1B are the cause of multiple endocrine neoplasia type 4 (MEN4) [MIM:610755]. Multiple endocrine neoplasia (MEN) syndromes are inherited cancer syndromes of the thyroid. MEN4 is a MEN-like syndrome with a phenotypic overlap of both MEN1 and MEN2.
- Sequence similaritiesBelongs to the CDI family.
- DomainA peptide sequence containing only AA 28-79 retains substantial Kip1 cyclin A/CDK2 inhibitory activity.
modificationsPhosphorylated; phosphorylation occurs on serine, threonine and tyrosine residues. Phosphorylation on Ser-10 is the major site of phosphorylation in resting cells, takes place at the G(0)-G(1) phase and leads to protein stability. Phosphorylation on other sites is greatly enhanced by mitogens, growth factors, cMYC and in certain cancer cell lines. The phosphorylated form found in the cytoplasm is inactivate. Phosphorylation on Thr-198 is required for interaction with 14-3-3 proteins. Phosphorylation on Thr-187, by CDK2 leads to protein ubiquitination and proteasomal degradation. Tyrosine phosphorylation promotes this process. Phosphorylation by PKB/AKT1 can be suppressed by LY294002, an inhibitor of the catalytic subunit of PI3K. Phosphorylation on Tyr-88 and Tyr-89 has no effect on binding CDK2, but is required for binding CDK4. Dephosphorylated on tyrosine residues by G-CSF.
Ubiquitinated; in the cytoplasm by the KPC complex (composed of RNF123/KPC1 and UBAC1/KPC2) and, in the nucleus, by SCF(SKP2). The latter requires prior phosphorylation on Thr-187. Ubiquitinated; by a TRIM21-containing SCF(SKP2)-like complex; leads to its degradation.
Subject to degradation in the lysosome. Interaction with SNX6 promotes lysosomal degradation.
- Cellular localizationNucleus. Cytoplasm. Endosome. Nuclear and cytoplasmic in quiescent cells. AKT-or RSK-mediated phosphorylation on Thr-198, binds 14-3-3, translocates to the cytoplasm and promotes cell cycle progression. Mitogen-activated UHMK1 phosphorylation on Ser-10 also results in translocation to the cytoplasm and cell cycle progression. Phosphorylation on Ser-10 facilitates nuclear export. Translocates to the nucleus on phosphorylation of Tyr-88 and Tyr-89. Colocalizes at the endosome with SNX6 and this leads to lysosomal degradation.
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Anti-p27 KIP 1 (phospho S10) antibody [EP233(2)Y] images
Immunohistochemistical detection of p27 KIP 1 (phospho S10) antibody [EP233(2)Y] (ab62364) on formaldehyde fixed paraffin-embedded monkey cerebellum sections. Antigen retrieval step: Heat mediated. Buffer: Citric acid solution. Permeabilization: None. Blocking step: 1% BSA for 10 mins. Primary antibody incubated at 1/1500 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody Name: anti rabbit IgG conjugated to biotin (1/200).
ICC/IF image of p27 KIP 1 (phospho S10) staining using ab62364 on Cor1 cells. Cells were fixed for 20 mins using 4% Formalin in PBS. After TBS washes, cells were then permeabilised/blocked simultaneously using antibody diluent before applying primary Ab.Some customers have sucessfully used the antibody in ICC/IF. Please see the abreview for the experimental conditions for the antibody staining.
All lanes : Anti-p27 KIP 1 (phospho S10) antibody [EP233(2)Y] (ab62364) at 1/10000 dilution
Lane 1 : MCF-7 cell lysate (untreated)
Lane 2 : MCF-7 cell lysate treated with AP
Lysates/proteins at 10 µg per lane.
goat anti-rabbit HRP labeled at 1/2000 dilution
Predicted band size : 22 kDa
Observed band size : 27 kDa (why is the actual band size different from the predicted?)
Immunohistochemistical detection of p27 KIP 1 (phospho S10) antibody [EP233(2)Y] (ab62364) on formaldehyde-fixed paraffin-embedded human tonsil sections. Antigen retrieval step: Heat mediated. Buffer Used: Citric acid. Permeabilization: None. Blocking step: 1% BSA for 10 mins @ 21°C. Primary antibody incubated at 1/500 for 2 hours @ 21°C in TBS/BSA/azide. Notes: A micropolymer HRP kit was used to obtain this intensity of immunostaining as I could not achieve a similar intensity of positivity on human tissues, under the same conditions (compare with results on rat/mouse/marmoset sections, using a std ABCpx detection system). There is one obvious metaphase nucleus in the Germinal centre of the submitted image: it is not positive.
ab62364 at 1/100 dilution staining p27 KIP 1 in human ovarian carcinoma by Immunohistochemistry, Paraffin-embedded tissue.
ab62364 at 1/100 dilution staining p27 KIP 1 in human tonsils by Immunohistochemistry, Paraffin-embedded tissue.
References for Anti-p27 KIP 1 (phospho S10) antibody [EP233(2)Y] (ab62364)
This product has been referenced in:
- Lin VT et al. TRIP6 regulates p27 KIP1 to promote tumorigenesis. Mol Cell Biol 33:1394-409 (2013). WB . Read more (PubMed: 23339869) »
- Uranbileg B et al. Cdc6 protein activates p27KIP1-bound Cdk2 protein only after the bound p27 protein undergoes C-terminal phosphorylation. J Biol Chem 287:6275-83 (2012). Read more (PubMed: 22223646) »