Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Rabbit polyclonal to p27 KIP 1 (phospho T157)
  • Suitable for: IHC-P, ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-p27 KIP 1 (phospho T157) antibody
    See all p27 KIP 1 primary antibodies

  • Description

    Rabbit polyclonal to p27 KIP 1 (phospho T157)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Cat, Dog, Pig

  • Immunogen

    Synthetic peptide corresponding to Human p27 KIP 1 aa 150 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab97604)

  • Positive control

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

  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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).

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

    • Western blot - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      Western blot - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      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
      All lanes : 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


      The 32kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Human Cyclin-dependent kinase inhibitor 1B (p27 KIP 1).
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      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
    • Immunocytochemistry/ Immunofluorescence - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      Immunocytochemistry/ Immunofluorescence - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      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.

    References (6)

    Publishing research using ab85047? Please let us know so that we can cite the reference in this datasheet.

    ab85047 has been referenced in 6 publications.

    • Chen Y  et al. Akt Regulated Phosphorylation of GSK-3ß/Cyclin D1, p21 and p27 Contributes to Cell Proliferation Through Cell Cycle Progression From G1 to S/G2M Phase in Low-Dose Arsenite Exposed HaCat Cells. Front Pharmacol 10:1176 (2019). PubMed: 31680960
    • Wu C  et al. SGK1 Governs the Reciprocal Development of Th17 and Regulatory T Cells. Cell Rep 22:653-665 (2018). ICC/IF ; Mouse . PubMed: 29346764
    • Taniuchi K  et al. WAVE2 is associated with poor prognosis in pancreatic cancers and promotes cell motility and invasiveness via binding to ACTN4. Cancer Med 7:5733-5751 (2018). PubMed: 30353690
    • Li Z  et al. Tumor-Secreted Exosomal miR-222 Promotes Tumor Progression via Regulating P27 Expression and Re-Localization in Pancreatic Cancer. Cell Physiol Biochem 51:610-629 (2018). PubMed: 30458449
    • Rostama B  et al. DLL4/Notch1 and BMP9 Interdependent Signaling Induces Human Endothelial Cell Quiescence via P27KIP1 and Thrombospondin-1. Arterioscler Thromb Vasc Biol 35:2626-37 (2015). PubMed: 26471266
    • 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 . PubMed: 23142123

    Images

    • Western blot - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      Western blot - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      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
      All lanes : 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


      The 32kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Human Cyclin-dependent kinase inhibitor 1B (p27 KIP 1).
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      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
    • Immunocytochemistry/ Immunofluorescence - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      Immunocytochemistry/ Immunofluorescence - Anti-p27 KIP 1 (phospho T157) antibody (ab85047)
      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.

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