712494

antibody from Invitrogen Antibodies

Targeting: BRPF1 BR140

Antibody data

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Validation data

BRPF1 protein structure - 712494 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of BRPF1 was achieved by transfecting HeLa cells with BRPF1 specific siRNA (Silencer® select Product # s15422 & s15423). Western blot analysis (Fig a) was performed using modified whole cell extracts (1% SDS) from BRPF1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-BRPF1 Recombinant Rabbit Polyclonal Antibody (Product # 712494, 1:250 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in the histogram (Fig b). Loss of signal upon siRNA mediated knockdown confirms that antibody is specific to BRPF1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-BRPF1 Recombinant Rabbit Polyclonal Antibody (Product # 712494) and 150, 140 kDa bands corresponding to isoforms of BRPF1 was observed across the cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), HT1080 (Lane 2), NTERA-2 (Lane 3) and Jurkat ( Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1:250 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).