PA1-26253

antibody from Invitrogen Antibodies

Targeting: CTSD CLN10, CPSD

Immunohistochemistry

Antibody data

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Validation data

CTSD protein structure - PA1-26253 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Cathepsin D was achieved by transfecting MCF7 with Cathepsin D specific siRNAs (Silencer® select Product # S137, S136). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Cathepsin D knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Cathepsin D Polyclonal Antibody (Product # PA1-26253, 1:1000 dilution ) 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 Cathepsin D.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Cathepsin D Polyclonal Antibody (Product # PA1-26253) and a 48 kDa band corresponding to Preprocathepsin D along with a 30 kDa band that corresponds to Cathepsin D heavy chain was observed across cell lines tested. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), Hep G2 (Lane 2), PANC-1 (Lane 3) and SK-BR-3 (Lane 4) were electrophoresed using NuPAGE™ 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 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).