Overview

  • Product nameAnti-p27 KIP 1 antibodySee all p27 KIP 1 primary antibodies ...
  • Description
    Rabbit polyclonal to p27 KIP 1
  • SpecificityReacts with p27 (also designated Kip1 p27)
  • Tested applicationsIP, IHC-P, WB, Sandwich ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    AGSVEQTPKKPGLRRRQT

    , corresponding to C terminal amino acids 181-198 of Human p27 KIP 1.

  • Positive control
    • Jurkat, HeLa, NIH/3T3, and KNRK

Properties

Applications

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.

Application Abreviews Notes
IP Use at an assay dependent dilution.
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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 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 Use a concentration of 1 µg/ml.

Target

  • 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.
  • Post-translational
    modifications
    Phosphorylated; 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.
  • Information by UniProt
  • Database links
  • Alternative names
    • AA408329 antibody
    • AI843786 antibody
    • Cdki1b antibody
    • CDKN 1B antibody
    • CDKN 4 antibody
    • CDKN1B antibody
    • CDKN4 antibody
    • CDN1B_HUMAN antibody
    • Cyclin Dependent Kinase Inhibitor 1B antibody
    • Cyclin dependent kinase inhibitor p27 antibody
    • Cyclin-dependent kinase inhibitor 1B (p27, Kip1) antibody
    • Cyclin-dependent kinase inhibitor 1B antibody
    • Cyclin-dependent kinase inhibitor p27 antibody
    • Cyclin-dependent kinase inhibitor p27 Kip1 antibody
    • KIP 1 antibody
    • KIP1 antibody
    • MEN1B antibody
    • MEN4 antibody
    • OTTHUMP00000195098 antibody
    • OTTHUMP00000195099 antibody
    • p27 antibody
    • p27 Kip1 antibody
    • P27-like cyclin-dependent kinase inhibitor antibody
    • P27KIP1 antibody
    see all

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.

    See Abreview

  • ICC/IF image of ab7961 stained Hek293 cells (ab7902). 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.

  • 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 (ab6721) was used as secondary antibody.

    See Abreview

  • 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.

    See Abreview

  • Standard curve for p27 KIP 1 (Analyte: ab56279); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal to p27 KIP 1 (ab54563) at 1µg/ml and Detector Antibody Rabbit polyclonal to p27 KIP 1 (ab7961) at 0.1µg/ml.

Protocols

References for Anti-p27 KIP 1 antibody (ab7961)

This product has been referenced in:
  • Yelamanchili SV  et al. The evolutionary young miR-1290 favors mitotic exit and differentiation of human neural progenitors through altering the cell cycle proteins. Cell Death Dis 5:e982 (2014). WB . Read more (PubMed: 24407235) »
  • Schimenti KJ  et al. AKAP9 is essential for spermatogenesis and sertoli cell maturation in mice. Genetics 194:447-57 (2013). Read more (PubMed: 23608191) »

See all 15 Publications for this product

Product Wall

Thank you for your enquiry regarding ab7961 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

Sample type: NCI-H460 - Recombinant protein
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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Cor1)
Specification Cor1
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 24 2011

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Monkey Tissue sections (Kidney)
Specification Kidney
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Dec 23 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Cerebellum)
Specification Cerebellum
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH6.3
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Nov 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (tonsil)
Specification tonsil
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate, 20 minutes, 100C
Permeabilization No
Username

Abcam user community

Verified customer

Submitted Nov 13 2009

I have received feedback from the laboratory and I would like to confirm that in our testing conditions the antibody detects a band at 27kda in HeLa lysate which is expected as the name "p27" suggest, therefore we think that the band around 390kda that...

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Thank you very much for your enquiry. I am sorry that this antibody did not work well with your tissue samples, but I am glad to hear that it worked in culture cells. It is good to hear that you are pleased with the products our company offers....

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Thank you for your enquiry. I noticed on your review that you were unhappy with the performance of ab7961. I would like to suggest the following modifications to your protocol to help you to get this antibody to work: Please try incubating wit...

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This antibody has not been tested on chicken so we can't guarantee that it will work. To the best of our knowledge chicken p27 has not yet been sequenced. The peptide sequence used to generate this antibody is: AGSVEQTPKKPGLRRRQT.

We suggest use the antibody at a starting dilution of 1:200 for IHC/ICC and adjust as needed.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"