PA5-17010

antibody from Invitrogen Antibodies

Targeting: UBE2N MGC8489, UBC13, UbcH-ben

Antibody data

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Validation data

UBE2N protein structure - PA5-17010 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of UBE2N was achieved by transfecting Hep G2 with UBE2N specific siRNAs (Silencer® select Product # s14596, s14595). Western blot analysis (Fig. a) was performed using whole cell extracts from the UBE2N knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with UBC13 Polyclonal Antibody (Product # PA5-17010, 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 UBE2N.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using anti-UBC13 Polyclonal Antibody (Product # PA5-17010) and a 17 kDa band corresponding to UBE2N was observed across all the cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HepG2 (Lane 1), HeLa (Lane 2), NIH:OVCAR-3 (Lane 3), BeWo (Lane 4), HL60 (Lane 5), MOLT4 (Lane 6), tissue extracts of Mouse Testis (Lane 7), Mouse Brain (Lane 8), Mouse Liver (Lane 9) and Mouse Ovary (Lane 10) 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).