MA5-14986

antibody from Invitrogen Antibodies

Targeting: CTNNA1 CAP102

Immunoprecipitation

Antibody data

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Validation data

CTNNA1 protein structure - MA5-14986 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Catenin alpha-1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR787281_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Catenin alpha-1 was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) andHeLa Catenin alpha-1 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-Catenin alpha-1 Monoclonal Antibody (J.783.6) (Product # MA5-14986, 1:1,000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Catenin alpha-1. An uncharacterized band was observed in all the samples at ~25 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Catenin alpha-1 Monoclonal Antibody (J.783.6)(Product # MA5-14986) and a 100kDa band corresponding to Catenin alpha-1 was observed across cell lines and tissue extracts tested except PC-3 and Daudi which is reported to be negative. A non-specific band around 28kDA has also been observed. Whole Cell Extract-WCL (30 µg lysate) of A-431 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3), LNCaP (Lane 4), PC-3 (Lane 5), Daudi (Lane 6), NIH/3T3 (Lane 7) and tissue extracts of Mouse Lung (Lane 8), Mouse Heart (Lane 9) and Rat Heart (Lane 10) were electrophoresed using NuPAGE™ 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 the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).