PA5-70526

antibody from Invitrogen Antibodies

Targeting: PHB2 Bap37, BCAP37, p22, REA

Antibody data

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Validation data

PHB2 protein structure - PA5-70526 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Prohibitin on mouse heart tissue lysate. The sample was probed with an REA polyclonal antibody (Product # PA5-70526) using a primary antibody dilution of 1.0 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Prohibitin2 was achieved by transfecting Hep G2 with Prohibitin2 specific siRNAs (Silencer® select Product # S22344, S225494). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Prohibitin2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Prohibitin Polyclonal Antibody (Product # PA5-70526, 1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Prohibitin2. Further, western blot was performed on Hep G2 cells transfected with Prohibitin1 specific siRNAs (Silencer® select Product # s10425, s10424). The blot, when probed with the same antibody, did not show any reduction in signal, concluding that the antibody is specific to Prohibitin2 alone, and not Prohibitin1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Prohibitin2 Polyclonal Antibody (Product # PA5-70526) and a 30 kDa band corresponding to Prohibitin2 , along with an uncharacterized band(*), was observed across cell lines and tissue tested. Whole cell extracts (30 µg lysate) of Caco-2 (Lane 1), Hep G2 (Lane 2), T-47D (Lane 3), A-431 (Lane 4), HeLa (Lane 5), NIH/3T3 (Lane 6) and PC-12 (Lane 7) and tissue extract of Mouse Heart (Lane 8) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). 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 µg/mL) 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).