Overview

  • Product nameAnti-Cdk4 antibodySee all Cdk4 primary antibodies ...
  • Description
    Rabbit polyclonal to Cdk4
  • SpecificityReacts with Cdk4
  • Tested applicationsICC/IF, IP, IHC-P, WB, Sandwich ELISA more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse Cdk4 (C terminal). The peptide used is within the 50 amino acids at the carboxy terminus of CDK4.
    (Peptide available as ab8010)

  • Positive control
    • HeLa and K-562

Properties

Applications

Our Abpromise guarantee covers the use of ab7955 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.Can be blocked with Cdk4 peptide (ab8010).

Starting dilution: 1:200

Sandwich ELISA Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [AF10A4] to Cdk4 (ab75511). For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [AF10A4] to Cdk4 (ab75511) as Capture.

Target

  • FunctionSer/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 diseaseDefects 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 similaritiesBelongs 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 localizationCytoplasm. 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.
  • Target information above from: UniProt accession P11802 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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

Anti-Cdk4 antibody images

  • ICC/IF image of ab7955 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 (ab7955, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) ab150077) 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.

  • Standard Curve for Cdk4 (Analyte: Cdk4 protein (T7 Tag) (ab40586)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [AF10A4] to Cdk4 (ab75511) at 5µg/ml and Detector Antibody Rabbit polyclonal to Cdk4 (ab7955) at 0.5µg/ml.
  • ab7955 staining Human placenta. Staining is localised to the cytoplasm.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

Protocols

References for Anti-Cdk4 antibody (ab7955)

This product has been referenced in:
  • Quinn J  et al. APRIL promotes cell-cycle progression in primary multiple myeloma cells: influence of D-type cyclin group and translocation status. Blood 117:890-901 (2011). Read more (PubMed: 20709908) »
  • Zeng X  et al. The Ras oncogene signals centrosome amplification in mammary epithelial cells through cyclin D1/Cdk4 and Nek2. Oncogene 29:5103-12 (2010). WB ; Human . Read more (PubMed: 20581865) »

See all 3 Publications for this product

Product Wall


We have not determined a concentration of ab7955 to use in IHC-P. This is up to the end user to determine depending on their experiment. We can however suggest to start at 5 ug/ml (i.e 1/40 dilution). Some of our customers have also had good resul...

Read More
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (tonsil, placenta)
Specification tonsil, placenta
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA, pH9, 20 min, 100C
Permeabilization No
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Submitted Nov 02 2011

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Colon tumor)
Specification Colon tumor
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: DAKO target retrieval solution
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Apr 23 2010

This antibody has been successfully used on formalin-fixed paraffin embedded normal human skin

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