- PA5-29247 - Invitrogen Antibodies CTNNA2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of alpha Catenin was performed by separating 30 µg of Whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a alpha Catenin Polyclonal Antibody (Product # PA5-29247) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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-alpha Catenin Polyclonal Antibody (Product # PA5-29247, 1:3,000 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 ~50 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using alpha Catenin Polyclonal Antibody (Product # PA5-29247). Sample (30 µg of whole cell lysate). Lane A: A431. Lane B: Hela . 7.5% SDS PAGE. Alpha Catenin Polyclonal Antibody (Product # PA5-29247) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using alpha Catenin Polyclonal Antibody (Product # PA5-29247). Sample (50 µg of whole cell lysate). Lane A: Mouse brain . 7.5% SDS PAGE. alpha Catenin Polyclonal Antibody (Product # PA5-29247) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-alpha Catenin Polyclonal Antibody(Product # PA5-29247) and a 100kDa band corresponding to alpha Catenin 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 45kDA was also observed across cell lines tested. Whole cell extracts (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) and Rat Heart (Lane 9) 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 (1:3000 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of alpha Catenin in methanol-fixed HeLa cells using an Alpha Catenin polyclonal antibody (Product # PA5-29247) at a 1:500 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of alpha Catenin was performed in paraffin-embedded mouse intestine tissue using Green: MAP2 Polyclonal Antibody (Product # PA5-85755) at a dilution of 1:200. Blue: Nuclear staining with Hoechst 33342. The signal was developed using a goat anti-rabbit IgG antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: alpha 1 Catenin antibody [N1N3] detects alpha 1 Catenin protein at membrane on mouse intestine by immunohistochemical analysis. Sample: Paraffin-embedded mouse intestine. Alpha 1 Catenin antibody [N1N3] (Product # PA5-29247) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: alpha 1 Catenin antibody [N1N3] detects alpha 1 Catenin protein at membrane on mouse prostate by immunohistochemical analysis. Sample: Paraffin-embedded mouse prostate. Alpha 1 Catenin antibody [N1N3] (Product # PA5-29247) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: alpha 1 Catenin antibody [N1N3] immunoprecipitates CTNNA1 protein in IP experiments. IP samples: HeLa whole cell extract. A. 30 µg HeLa whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of CTNNA1 protein by 4 µg alpha 1 Catenin antibody [N1N3] (Product # PA5-29247). 5 % SDS-PAGE. The immunoprecipitated CTNNA1 protein was detected by alpha 1 Catenin antibody [N1N3] (Product # PA5-29247) diluted at 1:500.

PA5-29247