MA5-14854

antibody from Invitrogen Antibodies

Targeting: AXIN1 PPP1R49

Immunoprecipitation

Antibody data

Submit

Validation data

AXIN1 protein structure - MA5-14854 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Axin1 was achieved by transfecting HEK-293 with Axin1 specific siRNAs (Silencer® select Product # s15814, s15815). Western blot analysis (Fig. a) was performed using whole cell extracts from the Axin1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Axin1 Monoclonal Antibody (Product # MA5-14854, 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 Axin1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Axin1 Monoclonal Antibody (Product # MA5-14854) and a 110 kDa band corresponding to Axin1 was observed across cell lines. Whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), NTERA-2 cl.D1 (Lane 2), U-937 (Lane 3), HEK-293 (Lane 4), HeLa (Lane 5), HCT 116 (Lane 6), U-87MG (Lane 7), A-431 (Lane 8), PC-12 (Lane 9) and tissue lysate of Mouse Brain (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).