Anti-CDK4 Antibody, Rabbit Polyclonal

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Anti-CDK4 Antibody, Rabbit Polyclonal General Information

Product name
Anti-CDK4 Antibody, Rabbit Polyclonal
Validated applications
WB,ELISA,IHC-P,ICC/IF,IP
Species reactivity
Reacts with: Human
Specificity
Human CDK4
Immunogen
Recombinant Human CDK4 Protein (Catalog#10732-H09B)
Preparation
Produced in rabbits immunized with purified, recombinant Human CDK4 (rh CDK4; Catalog#10732-H09B; NP_000066.1; Met1-Glu303). CDK4 specific IgG was purified by Human CDK4 affinity chromatography.
Source
Polyclonal Rabbit IgG
Purification
Protein A & Antigen Affinity
Formulation
0.2 μm filtered solution in PBS
Conjugate
Unconjugated
Form
Liquid
Shipping
This antibody is shipped as liquid solution at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
This antibody can be stored at 2℃-8℃ for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20℃ to -80℃. Preservative-Free. Avoid repeated freeze-thaw cycles.

Anti-CDK4 Antibody, Rabbit Polyclonal Validated Applications

Application Dilution
WB 1:500-1:2000
ELISA 1:5000-1:10000
IHC-P 1:500-1:2000
ICC-IF 1:300-1:10000
IP 1-4 μL/mg of lysate
Please Note: Optimal concentrations/dilutions should be determined by the end user.

Anti-CDK4 Antibody, Rabbit Polyclonal Images

Anti-CDK4 rabbit polyclonal antibody at 1:500 dilution

Lane A: Hela Whole Cell Lysate

Lane B: MCF7 Whole Cell Lysate

Lane C: K562 Whole Cell Lysate

Lysates/proteins at 30 μg per lane.

Secondary

Goat Anti- Rabbit  IgG H&L (Dylight 800)  at 1/10000 dilution.

Developed using the Odyssey technique.

Performed under reducing conditions.

Predicted band size:34 kDa

Observed band size:33 kDa

(We are unsure as to the identity of these extra bands.)

Immunochemical staining of human CDK4 in human breast carcinoma with rabbit polyclonal antibody (1:1000, formalin-fixed paraffin embedded sections).
Immunofluorescence staining of CDK4 in HeLa cells. Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS,blocked with 10% serum, and incubated with rabbit anti-human CDK4 polyclonal antibody (1:1000) at 4℃ overnight. Then cells were stained with the Alexa Fluor®488-conjugated Goat Anti-rabbit IgG secondary antibody (green) and counterstained with DAPI (blue). Positive staining was localized to cytoplasm and nucleus.

CDK4 was immunoprecipitated using:

Lane A:0.5 mg HepG2 Whole Cell Lysate

Lane B:0.5 mg k562 Whole Cell Lysate

Lane C:0.5 mg Hela Whole Cell Lysate

2 µL anti-CDK4 rabbit polyclonal antibody and 15 μl of 50 % Protein G agarose.

Primary antibody:

Anti-CDK4 rabbit polyclonal antibody,at 1:100 dilution

Secondary antibody:

Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5000 dilution

Developed using the odssey technique.

Performed under reducing conditions.

Predicted band size: 34 kDa

Observed band size: 34 kDa

Anti-CDK4 rabbit polyclonal antibody at 1:500 dilution

Lane A: CDK4 konckout Hela Whole Cell Lysate

Lane B: Hela Whole Cell Lysate

Lysates/proteins at 10 μg per lane.

Secondary

Goat Anti- Rabbit  IgG H&L (Dylight 800)  at 1/10000 dilution.

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size:34 kDa

Observed band size:33 kDa

(We are unsure as to the identity of these extra bands.)

Anti-CDK4 Antibody, Rabbit Polyclonal: Synonyms

Anti-CMM3 Antibody; Anti-PSK-J3 Antibody

CDK4 Background Information

CDK4 is a member of the Ser/Thr protein kinase family. It is highly similar to the gene products of S. cerevisiae cdc28 and S. pombe cdc2. It is a catalytic subunit of the protein kinase complex that is important for cell cycle G1 phase progression. The activity of CDK4 is restricted to the G1-S phase, which is controlled by the regulatory subunits D-type cyclins and CDK inhibitor p16(INK4a). CDK4 was shown to be responsible for the phosphorylation of retinoblastoma gene product. CDK4 is the 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. CDK4 has been shown to be mutated in some types of cancer, whilst a chromosomal rearrangement can lead to Cdk6 overexpression in lymphoma, leukemia and melanoma.
Full Name
cyclin-dependent kinase 4
Research Areas
Related Pathways
  • p53 Pathway
    p53 Pathway
References
  • Stepanova L, et al. (1996) Mammalian p50Cdc37 is a protein kinase-targeting subunit of Hsp90 that binds and stabilizes Cdk4. Genes Dev. 10(12):1491-502.
  • Lamphere L, et al. (1997) Interaction between Cdc37 and Cdk4 in human cells. Oncogene. 14(16): 1999-2004.
  • Dai K, et al. (1996) Physical interaction of mammalian CDC37 with CDK4. J Biol Chem. 271(36): 22030-4.
  • Sugimoto M, et al. (1999) Regulation of CDK4 activity by a novel CDK4-binding protein, p34SEI-1. Genes Dev. 13(22):3027-33.

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