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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
- Other assay [1]
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- Product number
- PA5-29930 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- gamma Catenin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: A431.
- Concentration
- 0.36 mg/mL
Submitted references Central role for GSK3β in the pathogenesis of arrhythmogenic cardiomyopathy.
Chelko SP, Asimaki A, Andersen P, Bedja D, Amat-Alarcon N, DeMazumder D, Jasti R, MacRae CA, Leber R, Kleber AG, Saffitz JE, Judge DP
JCI insight 2016 Apr 21;1(5)
JCI insight 2016 Apr 21;1(5)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using gamma Catenin Polyclonal Antibody (Product # PA5-29930). Sample (30 µg of whole cell lysate). Lane A: A431 . 7.5% SDS PAGE. JUP antibody . gamma Catenin Polyclonal Antibody (Product # PA5-29930) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of gamma Catenin was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR680765_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of gamma Catenin was performed by loading 30 µg of A-431 Wild type (Lane 1), A-431 Cas9 (Lane 2) andA-431 gamma Catenin KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with gamma Catenin Polyclonal Antibody (Product # PA5-29930, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 dilution) using the iBright™ FL 1500 (Product # A44115). 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 gamma Catenin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-gamma Catenin Polyclonal Antibody(Product # PA5-29930) and a 82kDa band corresponding to gamma Catenin was observed across the cell lines tested except THP-1 and Daudi. Whole cell extracts (30 µg lysate) of HaCaT (Lane 1), Caco-2 (Lane 2), A-431 (Lane 3), THP-1 (Lane 4) and Daudi (Lane 5) electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of gamma Catenin was performed using 70% confluent log phase HaCaT cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with gamma Catenin Polyclonal Antibody (Product # PA5-29930) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing membrane and cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- gamma Catenin Polyclonal Antibody detects gamma Catenin protein at cytosol and membrane on human colon carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human colon carcinoma. Gamma Catenin Polyclonal Antibody (Product # PA5-29930) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- gamma Catenin Polyclonal Antibody detects gamma Catenin protein at cytosol and membrane on mouse esophagus by immunohistochemical analysis. Sample: Paraffin-embedded mouse esophagus. Gamma Catenin Polyclonal Antibody (Product # PA5-29930) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Gamma Catenin antibody immunoprecipitates Gamma Catenin protein in IP experiments. IP Sample: A431 whole cell lysate/extract A : 30 µg whole cell lysate/extract of Gamma Catenin protein expressing A431 cells B : Control with 2.5 µg of pre-immune rabbit IgG C : Immunoprecipitation of Gamma Catenin by 2.5 µg of Gamma Catenin antibody (Product # PA5-29930) 7.5% SDS-PAGE The immunoprecipitated Gamma Catenin protein was detected by Gamma Catenin antibody (Product # PA5-29930) diluted at 1:1,000. Anti-rabbit IgG (HRP) was used as a secondary reagent.
