PA5-17053

antibody from Invitrogen Antibodies

Targeting: HSPA4 HS24/P52, HSPH2

Antibody data

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Validation data

HSPA4 protein structure - PA5-17053 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of HSPA4 was achieved by transfecting Hep G2 with HSPA4 specific siRNAs (Silencer® select Product # s6976, s6978). Western blot analysis (Fig. a) was performed using whole cell extracts from the HSPA4 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HSPA4 Polyclonal Antibody (Product # PA5-17053, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25µg/ml, 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 HSPA4.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-HSPA4 Rabbit Polyclonal Antibody (Product # PA5-17053) and a 110kDa band corresponding to HSPA4 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), HEK-293 (Lane 3), A549 (Lane 4), COS-7 (Lane 5), Mouse Testis (Lane 6) and Rat Ovary (Lane 7) 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 (1:1000 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).