Recombinant
RabMAb

Recombinant Anti-p27 KIP 1 antibody [EPR18388-138] - BSA and Azide free (ab227911)

Overview

  • Product name

    Anti-p27 KIP 1 antibody [EPR18388-138] - BSA and Azide free
    See all p27 KIP 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR18388-138] to p27 KIP 1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P46414

  • Positive control

    • IHC: Mouse testis tissue.
  • General notes

    Ab227911 is the carrier-free version of ab190851. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab227911 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab227911 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 Use at an assay dependent concentration. Detects a band of approximately 18, 27 kDa (predicted molecular weight: 22 kDa).
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

Target

  • Function

    Important 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 specificity

    Expressed in all tissues tested. Highest levels in skeletal muscle, lowest in liver and kidney.
  • Involvement in disease

    Defects 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 similarities

    Belongs to the CDI family.
  • Domain

    A 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 localization

    Nucleus. 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

Images

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling p27 KIP 1 with ab190851 at 1/40000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Nuclear staining on Sertoli cells and Leydig cells of rat testis is observed (PMID: 10098522). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190851).

  • p27 KIP 1 was immunoprecipitated from 0.35 mg of C2C12 (mouse myoblast cell line) whole cell lysate with ab190851 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab190851 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as for detection at 1/10000 dilution.

    Lane 1: C2C12 whole cell lysate 10 µg (Input). 

    Lane 2: ab190851 IP in C2C12 whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab190851 in C2C12 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190851).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling p27 KIP 1 with ab190851 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mainly nuclear staining in the NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190851).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling p27 KIP 1 with ab190851 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mainly nuclear staining in the Neuro-2a cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190851).

  • Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling p27 KIP 1 with ab190851 at 1/40000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Nuclear staining in the rat cerebral cortex is observed (PMID: 19852587). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190851).

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling p27 KIP 1 with ab190851 at 1/40000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Nuclear staining on mouse kidney is observed (PMID: 26823281). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190851).

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling p27 KIP 1 with ab190851 at 1/40000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Nuclear staining on Leydig cells and Sertoli cells of mouse testis is observed (PMID: 10098522). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190851).

References

ab227911 has not yet been referenced specifically in any publications.

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