Overview

  • Product nameAnti-p27 KIP 1 antibody [Y236]See all p27 KIP 1 primary antibodies ...
  • Description
    Rabbit monoclonal [Y236] to p27 KIP 1
  • Specificityab32034 recognises p27(Kip1).
  • Tested applicationsIHC-Fr, IHC-P, ICC, IHC-FrFl, WB, ICC/IF, Flow Cyt, IP more details
  • Species reactivity
    Reacts with: Rat, Human

    Does not react with

    Mouse
  • Immunogen

    A synthetic peptide corresponding to residues in the C-term of human p27(Kip1) was used as immunogen

  • Positive control
    • MCF-7 cell lysate, PC-12 cells, human breast carcinoma
  • General notes

    Produced under U.S. Patent No. 5,675,063.

     

    A trial size is available for this product.

Properties

Applications

Our Abpromise guarantee covers the use of ab32034 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-FrFl Use at an assay dependent concentration. PubMed: 23880266
WB 1/1000. Detects a band of approximately 27 kDa (predicted molecular weight: 22 kDa).
ICC/IF 1/100 - 1/250.
Flow Cyt 1/40.
IP 1/50.

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.
  • Target information above from: UniProt accession P46527 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • AA408329 antibody
    • AI843786 antibody
    • Cdki1b antibody
    • CDKN 1B antibody
    • CDKN 4 antibody
    • CDKN1B antibody
    • CDKN1B antibody
    • CDKN4 antibody
    • CDKN4 antibody
    • CDN1B_HUMAN antibody
    • Cyclin Dependent Kinase Inhibitor 1B antibody
    • Cyclin Dependent Kinase Inhibitor 1B antibody
    • Cyclin dependent kinase inhibitor p27 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 [Y236] images

  • Anti-p27 KIP 1 antibody [Y236] (ab32034) at 1/1000 dilution + MCF-7 cell lysate

    Predicted band size : 22 kDa
    Observed band size : 27 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma
  • Immunofluorescent staining of PC-12 cells
  • ab32034 staining p27 KIP1 in MCF7 cells treated with NS 398 (ab120295), by ICC/IF. Increase in p27 KIP1 expression correlates with increased concentration of NS 398, as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab120295 (NS 398) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32034 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.
  • ab32034 staining p27 KIP 1 in human stomach tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with acetone and then blocked with 5% serum for 1 hour at 23°C followed by incubation with the primary antibody, at a 1/100 dilution, for 1 hour at 23°C. An undiluted HRP-conjugated goat polyclonal was used as secondary antibody.

    See Abreview

  • ab32034 showing positive staining in Colonic adenocarcinoma tissue.

  • ab32034 showing positive staining in Glioma tissue.

  • ab32034 showing positive staining in Ovarian carcinoma tissue.

  • ab32034 showing positive staining in Stomach adenocarcinoma tissue.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-p27 KIP 1 antibody [Y236] (ab32034)

This product has been referenced in:
  • Klinz FJ  et al. Nuclear expression of p27(Kip1) is associated with in vivo differentiation of adult human odontoblasts. J Endod 39:995-1000 (2013). IHC-FrFl ; Human . Read more (PubMed: 23880266) »
  • Zhu Y  et al. Involvement of decreased hypoxia-inducible factor 1 activity and resultant G(1)-S cell cycle transition in radioresistance of perinecrotic tumor cells. Oncogene : (2012). Flow Cyt, WB ; Human . Read more (PubMed: 22710721) »

See all 5 Publications for this product

Product Wall

Application Western blot
Loading amount 50000 cells
Gel Running Conditions Reduced Denaturing
Sample Rat Cell lysate - whole cell (rat vascula smooth muscle cells)
Specification rat vascula smooth muscle cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted Aug 22 2013

Application Western blot
Sample Human Cell lysate - whole cell (293)
Loading amount 20 µg
Specification 293
Treatment 10 uM MG132 4 hours
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted May 13 2013

Application Immunocytochemistry
Sample Human Cultured Cells (AGS Gastric Carcinoma cells)
Specification AGS Gastric Carcinoma cells
Fixative Formaldehyde
Permeabilization Yes - 0.025% Triton-X in TBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted May 19 2010

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (AGS)
Specification AGS
Fixative Formaldehyde
Permeabilization Yes - 0.025% Triton-X in TBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted May 12 2010

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Stomach)
Specification Stomach
Fixative Acetone
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Apr 01 2010

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Stomach)
Specification Stomach
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Mar 31 2010

Application Flow Cytometry
Sample Human Cell (AGS human gastric carcinoma cell line)
Specification AGS human gastric carcinoma cell line
Preparation Cell harvesting/tissue preparation method: Accutase/centrifugation
Sample buffer: PBS 5%FCS
Fixation Formaldehyde
Permeabilization Yes - 0.025% Triton-X in PBS
Gating Strategy Size gated AGS
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Submitted Feb 11 2010

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (AGS)
Total protein in input 50 µg
Specification AGS
Immuno-precipitation step Protein A/G
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Submitted Feb 01 2010

Application Western blot
Sample Human Cell lysate - whole cell (AGS cells)
Loading amount 20 µg
Specification AGS cells
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Sep 02 2009

Application Western blot
Sample Human Cell lysate - whole cell (muscle)
Loading amount 25 µg
Specification muscle
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Feb 01 2008

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