Overview

  • Product name
    Anti-p27 KIP 1 antibody [SX53G8]
    See all p27 KIP 1 primary antibodies
  • Description
    Mouse monoclonal [SX53G8] to p27 KIP 1
  • Host species
    Mouse
  • Specificity
    ab193379 is highly specific and shows no cross-reaction with other related mitotic inhibitors.
  • Tested applications
    Suitable for: Flow Cyt, IP, ICC/IF, IHC-Fr, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Monkey
    Predicted to work with: Cat, Dog, Chinese hamster
  • Immunogen

    Recombinant full length protein (proprietary-tag) corresponding to Human p27 KIP 1 aa 1-198.
    Sequence:

    MSNVRVSNGSPSLERMDARQAEHPKPSACRNLFGPVDHEELTRDLEKHCR DMEEASQRKWNFDFQNHKPLEGKYEWQEVEKGSLPEFYYRPPRPPKGACK VPAQESQDVSGSRPAAPLIGAPANSEDTHLVDPKTDPSDSQTGLAEQCAG IRKRPATDDSSTQNKRANRTEENVSDGSPNAGSVEQTPKKPGLRRRQT


    Database link: P46527

  • Positive control
    • WB: HAP1, HeLa and MCF7 cell lysate. IHC: Human colon tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab193379 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
Flow Cyt Use 0.5-1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

IP Use at 1-2 µg/mg of lysate.
ICC/IF Use a concentration of 0.5 - 1 µg/ml.
IHC-Fr Use a concentration of 0.5 - 1 µg/ml.
WB Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 22 kDa.
IHC-P Use a concentration of 0.5 - 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: CDKN1B knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: MCF7 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab193379 observed at 30 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab193379 was shown to specifically recognize CDKN1B in wild-type HAP1 cells as well as additional cross-reactive bands. No bands were observed when CDKN1B knockout samples were used. Wild-type and CDKN1B knockout samples were subjected to SDS-PAGE.  ab193379 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 µg/mL and 1/10,000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

     

  • Ab193379 staining p27 KIP1 in HeLa cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/200 in PBS) for 1 hour at 22°C. A Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) was used as the secondary antibody.

    See Abreview

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human colon tissue labeling p27 KIP 1 with ab193379 at 1 µg/mL.

References

This product has been referenced in:
  • Gong SJ  et al. Upregulation of PP2Ac predicts poor prognosis and contributes to aggressiveness in hepatocellular carcinoma. Cancer Biol Ther 17:151-62 (2016). Read more (PubMed: 26618405) »

See 1 Publication for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X-100
Specification
HeLa
Fixative
Paraformaldehyde
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Jul 26 2017

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse breast cancer cell line)
Gel Running Conditions
Reduced Denaturing (Gel 15%)
Loading amount
30 µg
Specification
Mouse breast cancer cell line
Blocking step
Odyssey blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 07 2017

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

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