Anti-Cdk4 antibody (ab95255)

Overview

• Product nameAnti-Cdk4 antibodySee all Cdk4 primary antibodies ...
• Description
  Rabbit polyclonal to Cdk4
• Tested applicationsWB, ICC/IF more details
• Species reactivity
  Reacts with: Human
  Predicted to work with: Pig
• Immunogen

  Synthetic peptide conjugated to KLH derived from within residues 250 to the C-terminus of Human Cdk4.

  Read Abcam's proprietary immunogen policy

  (Peptide available as ab109075.)

• Positive controlThis antibody gave a positive signal in both MCF7 and HeLa whole cell lysates.

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: 1% BSA, PBS, pH 7.4
• Concentration information loading...
• PurityImmunogen affinity purified
• Clonality Polyclonal
• IsotypeIgG
• Research Areas
  • Cancer
  • Cell cycle
  • Kinases/phosphatases
  • Cdks

  • Epigenetics and Nuclear Signaling
  • Cell cycle
  • Kinases/Phosphatases
  • Cdks

  • Cell Biology
  • Cell Cycle
  • Kinases/Phosphatases
  • Cdks

Applications

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
  modificationsPhosphorylation 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
  • Entrez Gene: 1019 Human
  • Entrez Gene: 100144492 Pig
  • Omim: 123829 Human
  • SwissProt: P11802 Human
  • SwissProt: P79432 Pig
  • Unigene: 95577 Human

• Alternative names
  Cdk 4 antibodycdk4 antibodyCDK4 protein antibody

  CDK4_HUMAN antibodyCell division kinase 4 antibodyCell division protein kinase 4 antibodyCMM 3 antibodyCMM3 antibodyCrk3 antibodyCyclin dependent kinase 4 antibodyCyclin-dependent kinase 4 antibodyMelanoma cutaneous malignant 3 antibodyMGC14458 antibodyp34 cdk4 antibodyPSK J3 antibodyPSK-J3 antibody

  see all

Anti-Cdk4 antibody images

• 

  Western blot - Cdk4 antibody (ab95255)
  Anti-Cdk4 antibody (ab95255) at 1 µg/ml + MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
  
  Secondary
  Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 33 kDa
  Observed band size : 33 kDa
  Additional bands at : 110 kDa,16 kDa. We are unsure as to the identity of these extra bands.
  
  Exposure time : 20 minutes
• 

  Immunocytochemistry/ Immunofluorescence - Cdk4 antibody (ab95255)
  ICC/IF image of ab95255 stained MCF7 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 (ab95255, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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 and HepG2 cells at 5µg/ml, and in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 5µg/ml.