Overview

  • Product nameAnti-p27 KIP 1 (phospho T157) antibody
    See all p27 KIP 1 primary antibodies
  • Description
    Rabbit polyclonal to p27 KIP 1 (phospho T157)
  • Tested applicationsIHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Cat, Dog, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human p27 KIP 1.

    (Peptide available as ab97604.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; Caco2; MCF7.

Properties

Applications

Our Abpromise guarantee covers the use of ab85047 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 22 kDa).Can be blocked with Human p27 KIP 1 (phospho T157) peptide (ab97604).

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 T157) antibody images

  • All lanes : Anti-p27 KIP 1 (phospho T157) antibody (ab85047) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
    Lane 3 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human p27 KIP 1 (phospho T157) peptide (ab97604) at 1 µg/ml
    Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human p27 KIP 1 (phospho T157) peptide (ab97604) at 1 µg/ml
    Lane 7 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate with Human p27 KIP 1 (phospho T157) peptide (ab97604) at 1 µg/ml
    Lane 8 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with Human p27 KIP 1 (phospho T157) peptide (ab97604) at 1 µg/ml
    Lane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with p27 KIP 1 peptide (ab116623) at 1 µg/ml
    Lane 10 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with p27 KIP 1 peptide (ab116623) at 1 µg/ml
    Lane 11 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate with p27 KIP 1 peptide (ab116623) at 1 µg/ml
    Lane 12 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with p27 KIP 1 peptide (ab116623) at 1 µg/ml

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 22 kDa
    Observed band size : 32 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa,75 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes
  • ICC/IF image of ab85047 stained HeLa cells. The cells were 4% PFA 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 (ab85047, 5µg/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. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml.
  • IHC image of p27 KIP 1 (phospho T157) staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85047, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX

References for Anti-p27 KIP 1 (phospho T157) antibody (ab85047)

This product has been referenced in:
  • Inge LJ  et al. Dasatinib, a small molecule inhibitor of the Src kinase, reduces the growth and activates apoptosis in pre-neoplastic Barrett's esophagus cell lines: evidence for a noninvasive treatment of high-grade dysplasia. J Thorac Cardiovasc Surg 145:531-8 (2013). Human . Read more (PubMed: 23142123) »

See 1 Publication for this product

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