Anti-p27 KIP 1 antibody (ab7961)
- Product nameAnti-p27 KIP 1 antibodySee all p27 KIP 1 primary antibodies ...
- DescriptionRabbit polyclonal to p27 KIP 1
- SpecificityReacts with p27 (also designated Kip1 p27)
- Tested applicationsIP, IHC-P, WB, Sandwich ELISA, ICC/IF more details
- Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide: AGSVEQTPKKPGLRRRQT, corresponding to C terminal amino acids 181-198 of Human p27 KIP 1.
- Positive control
- Jurkat, HeLa, NIH/3T3, and KNRK
- Storage instructionsStore at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
- Storage bufferPBS with 0.1% sodium azide
- Concentration information loading...
- PurityIgG fraction
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab7961 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||IP: Use at an assay dependent dilution.|
|IHC-P||IHC-P: 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||WB: Use at an assay dependent dilution. Detects a band of approximately 27 kDa (predicted molecular weight: 22 kDa).Can be blocked with p27 KIP 1 peptide (ab8016).|
|Sandwich ELISA||sELISA: Use a concentration of 0.1 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal to p27 KIP 1 (ab54563). For sandwich ELISA, use this antibody as Detection at 0.1µg/ml with Ab54563 as Capture.|
|ICC/IF||ICC/IF: Use a concentration of 1 µg/ml.|
- 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 antibody images
ICC/IF Image of p27 KIP1 staining in Mouse Cor1 cells with ab7961. The cells were permeabilised with 0.1% Triton. Cells were blocked with 1% BSA for 20 minutes at 21°C and incubated with ab7961 (1/250 dilution) for 2 hours at 21°C.
ICC/IF image of ab7961 stained Hek293 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7961, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistical detection of p27 KIP 1 antibody (ab7961) on formaldehyde-fixed paraffin-embedded mouse cerebellum sections. Antigen retrieval step: Heat mediated. Buffer Used: Citric acid pH6.3. Blocking step: 1% BSA for 10 mins @ 21°C. Primary antibody: 1/250 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody: anti rabbit IgG Conjugation: Biotin. An excellent immunostaining pattern that is most evident in nuclei. However, a significant number of nuclei are negative for this Ab and the significance of this is as yet unknown.
ab7961 staining 927 KIP1 in human tonsillar tissue sections by IHC-P (formaldehyde-fixed paraffin-embedded sections). Tissue samples were fixed with formaldehyde. Antigen retrieval was by heat mediation in citrate buffer (20 minutes at 100°C). Samples were incubated with primary antibody 1/25 (background reducer diluent) for 20 minutes at 25°C. An undiluted HRP polymer Goat anti mouse/rabbit IgG was used as secondary antibody.
Anti-p27 KIP 1 antibody (ab7961) at 1 µg/ml +
Human p27 KIP 1 full length protein (ab56279) at 0.01 µg
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 10 seconds
References for Anti-p27 KIP 1 antibody (ab7961)
This product has been referenced in:
- Schimenti KJ et al. AKAP9 is essential for spermatogenesis and sertoli cell maturation in mice. Genetics 194:447-57 (2013). Read more (PubMed: 23608191) »
- Li YG et al. Polysaccharide from Phellinus linteus induces S-phase arrest in HepG2 cells by decreasing calreticulin expression and activating the P27kip1-cyclin A/D1/E-CDK2 pathway. J Ethnopharmacol 150:187-95 (2013). Read more (PubMed: 24001891) »