MA5-15119

antibody from Invitrogen Antibodies

Targeting: JUN AP-1, c-Jun

Antibody data

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Validation data

JUN protein structure - MA5-15119 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of c-Jun was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR905187_SGM) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of JUN was performed by loading 30 µg of HEK-293 wild type (Lane-1), HEK-293 Cas9 (Lane-2) and HEK-293 c-Jun KO (Lane-3) modified 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 c-Jun Monoclonal Antibody (T.731.6) (Product # MA5-15119, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10,000 dilution) using the iBright™ FL1500 (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to c-Jun.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of c-Jun in extracts from 293 cells, untreated or UV-treated, using c-Jun monoclonal antibody (Product # MA5-15119).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-c-Jun Monoclonal Antibody (T.731.6) (Product # MA5-15119) and a 40 kDa band corresponding to Transcription factor AP-1 was observed across tested cell lines. Nuclear enriched extracts (40 µg lysate) of A-431 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3), HEK-293 (Lane 4), NIH/3T3 (Lane 5) were electrophoresed using NuPAGE™ 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 the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).