MA1-12299

antibody from Invitrogen Antibodies

Targeting: CDK4 PSK-J3

Immunoprecipitation

Antibody data

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Validation data

CDK4 protein structure - MA1-12299 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of CDK4 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1007917_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of CDK4 was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) andHeLa CDK4 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-CDK4 Monoclonal Antibody (DCS31.2) (Product # MA1-12299, 1:1,000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to CDK4.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-CDK4 Monoclonal Antibody (DCS31.2) (Product # MA1-12299) and a 33 kDa band corresponding to CDK4 was observed across the cell lines tested and tissues tested. Circulating tissue IgG band (*) at ~ 25 kDa was observed in Mouse Lung tissue. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), K-562 (Lane 2), A549 (Lane 3), HeLa (Lane 4), NIH/3T3 (Lane 5), PC12 (Lane 6), Mouse Lung (Lane 7), Rat Lung (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1,000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4,000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).