Overview

  • Product name

  • Description

    Rabbit polyclonal to Cdk4
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Pig
  • Immunogen

    Recombinant fragment, corresponding to a region within amino acids 148-303 of Human Cdk4 (Uniprot ID: P11802).

  • Positive control

    • Recombinant Human Cdk4 protein (ab126909) can be used as a positive control in WB. 293T, H1299, HeLa and NIH 3T3 whle cell lysates; paraformaldehyde-fixed HeLa cells; Human gastric carcinoma tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab137675 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 1/500 - 1/3000. Predicted molecular weight: 34 kDa.
ICC/IF 1/100 - 1/1000.
IHC-P 1/100 - 1/1000.

Target

  • Function

    Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex.
  • Involvement in disease

    Defects in CDK4 are a cause of susceptibility to cutaneous malignant melanoma type 3 (CMM3) [MIM:609048]. Malignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites.
  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Phosphorylation at Thr-172 is required for enzymatic activity. Phosphorylated, in vitro, at this site by CCNH-CDK7, but, in vivo, appears to be phosphorylated by a proline-directed kinase. In the cyclin D-CDK4-CDKN1B complex, this phosphorylation and consequent CDK4 enzyme activity, is dependent on the tyrosine phosphorylation state of CDKN1B. Thus, in proliferating cells, CDK4 within the complex is phosphorylated on Thr-172 in the T-loop. In resting cells, phosphorylation on Thr-172 is prevented by the non-tyrosine-phosphorylated form of CDKN1B.
  • Cellular localization

    Cytoplasm. Nucleus. Membrane. Cytoplasmic when non-complexed. Forms a cyclin D-CDK4 complex in the cytoplasm as cells progress through G(1) phase. The complex accumulates on the nuclear membrane and enters the nucleus on transition from G(1) to S phase. Also present in nucleoli and heterochromatin lumps. Colocalizes with RB1 after release into the nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Cdk 4 antibody
    • cdk4 antibody
    • CDK4 protein antibody
    • CDK4_HUMAN antibody
    • Cell division kinase 4 antibody
    • Cell division protein kinase 4 antibody
    • CMM 3 antibody
    • CMM3 antibody
    • Crk3 antibody
    • Cyclin dependent kinase 4 antibody
    • Cyclin-dependent kinase 4 antibody
    • Melanoma cutaneous malignant 3 antibody
    • MGC14458 antibody
    • p34 cdk4 antibody
    • PSK J3 antibody
    • PSK-J3 antibody
    see all

Images

  • All lanes : Anti-Cdk4 antibody (ab137675) at 1/1000 dilution

    Lane 1 : Epithelial OVCAR whole cell lysate - untreated
    Lane 2 : Epithelial OVCAR whole cell lysate treated with 100µM Perifosine for 24 hours
    Lane 3 : Epithelial OVCAR whole cell lysate treated with 200µM Perifosine for 24 hours

    Lysates/proteins at 100 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 34 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 minutes


    10% SDS-PAGE.

    Blocked with 5% milk for 12 hours at 4°C.

    Incubated with the primary antibody in 2% milk for 12 hours at 4°C.

    See Abreview

  • Immunofluorescent analysis of paraformaldehyde-fixed HeLa cells labelling Cdk4 with ab137675 at 1/500 dilution. Lower image shows cells co-stained with Hoechst 33342.
  • Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue, labelling Cdk4 using ab137675 at 1/500 dilution.
  • Anti-Cdk4 antibody (ab137675) at 1/1000 dilution + 293T whole cell lysate at 30 µg

    Predicted band size: 34 kDa



    12% SDS PAGE
  • Anti-Cdk4 antibody (ab137675) at 1/1000 dilution + NIH 3T3 whole cell lysate at 30 µg

    Predicted band size: 34 kDa



    12% SDS PAGE

References

This product has been referenced in:

  • Song J  et al. LATS2 overexpression attenuates the therapeutic resistance of liver cancer HepG2 cells to sorafenib-mediated death via inhibiting the AMPK-Mfn2 signaling pathway. Cancer Cell Int 19:60 (2019). Read more (PubMed: 30923462) »
  • Tormo E  et al. The miRNA-449 family mediates doxorubicin resistance in triple-negative breast cancer by regulating cell cycle factors. Sci Rep 9:5316 (2019). Read more (PubMed: 30926829) »
See all 20 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (Epithelial OVCAR cell line)
Gel Running Conditions
Reduced Denaturing (10 % SDS-PAGE)
Loading amount
100 µg
Treatment
Perifosine 24 hr
Specification
Epithelial OVCAR cell line
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

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Verified customer

Submitted Feb 23 2017

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