Cdk4 Antibody (YA513)

Cdk4 Antibody (YA513) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Cdk4.

CDK4 protein serves as the Ser/Thr-kinase component within cyclin D-CDK4 (DC) complexes, orchestrating the phosphorylation and inhibition of members belonging to the retinoblastoma (RB) protein family, including RB1. This regulatory activity is pivotal in controlling the cell cycle during the G(1)/S transition. The phosphorylation of RB1 instigates the dissociation of the transcription factor E2F from the RB/E2F complexes, facilitating the subsequent transcription of E2F target genes that drive progression through the G(1) phase. Particularly notable is the hypophosphorylation of RB1 occurring in early G(1) phase. As essential integrators of diverse mitogenic and antimitogenic signals, cyclin D-CDK4 complexes play a central role in cell cycle regulation. Additionally, CDK4 protein exhibits the capability to phosphorylate SMAD3 in a cell-cycle-dependent manner, thereby repressing its transcriptional activity. CDK4 is a crucial component of the ternary complex, cyclin D/CDK4/CDKN1B, which is indispensable for the nuclear translocation and activity of the cyclin D-CDK4 complex.

Free

P30285

Predicted band size: 34 kDa; Observed band size: 30 kDa

Protein A affinity purified.

Cytoplasm, Nucleus, Membrane.

Non-conjugated

Unmodified

AB_3102068

Cell Biology

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:1,000-1:2,000; ICC/IF: 1:50-1:200; IHC-P: 1:1,000; IP: 1:50

• 
  Western blot analysis of extracts from Hela (lane 2(20μg), NIH/3T3 (lane 3(20μg), C2C12 (lane 4(20μg) using Integrin alpha 5 Antibody (HY-P80077) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80416, 1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of Hela cells labeling Cdk4 with Cdk4antibody (HY-P80077) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Cdk4 antibody (HY-P80077) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse colon tissue using Cdk4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80077, 1/500) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunocytochemistry analysis of NIH3T3 cells labeling Cdk4 with Cdk4 antibody (HY-P80077) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Cdk4 antibody (HY-P80077) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse colon tissue using Cdk4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80077, 1/500) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, ICC/IF, IHC-P, IP

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Mouse Cdk4.AA range:180-303.