RabMAb

Anti-p27 KIP 1 (phospho S10) antibody [EP233(2)Y] (ab62364)

Overview

  • Product nameAnti-p27 KIP 1 (phospho S10) antibody [EP233(2)Y]See all p27 KIP 1 primary antibodies ...
  • Description
    Rabbit monoclonal [EP233(2)Y] to p27 KIP 1 (phospho S10)
  • Specificityab62364 detects p27 KIP 1 phosphorylated at Serine 10.
  • Tested applicationsWB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Monkey
  • Immunogen

    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 notes

    Produced under U.S. Patent No. 5,675,063. A 40 μl trial size is available to purchase for this antibody.

Properties

Applications

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.

Application Abreviews Notes
WB 1/10000 - 1/20000. Detects a band of approximately 27 kDa (predicted molecular weight: 22 kDa).
IP 1/30.
IHC-P 1/200 - 1/1500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notesIs unsuitable for Flow Cyt or ICC.
  • 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 (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).

      See Abreview

    • 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.

      See Abreview

    • 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.

      Secondary
      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.

      See Abreview

    • 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) »

    See all 7 Publications for this product

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    Read More
    Application Immunocytochemistry/ Immunofluorescence
    Sample Mouse Cell (Cor1 cells)
    Specification Cor1 cells
    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 Mar 28 2011

    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
    Permeabilization No
    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

    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: Citric acid
    Permeabilization No
    Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Nov 02 2010

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

    Mr. Carl Hobbs

    Verified customer

    Submitted Nov 02 2010

    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: EDTA, 20 minutes,100oC
    Permeabilization No
    Username

    Abcam user community

    Verified customer

    Submitted Nov 13 2009

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