701857

antibody from Invitrogen Antibodies

Targeting: WNT4 WNT-4

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

701857

Provider

Invitrogen Antibodies

Product name

WNT4 Recombinant Rabbit Monoclonal Antibody (9H2L10)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Pig, Rat and Mouse.

Antibody clone number

9H2L10

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF-7 (Lane1), A-431 (Lane 2), HT-29 (Lane 3), A549 (Lane 4), K-562 (lane 5), MDA-MB-231 (Lane 6), Colo 205 (Lane 7), HeLa (Lane 8), Caco2 (Lane 9) and Jurkat (Lane 10). The blots were probed with WNT4 Recombinant Rabbit Monoclonal Antibody (Product # 701857, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 40 kDa band corresponding to WNT4 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of WNT4 was achieved by transfecting A-431 cells with WNT4 specific validated siRNAs (Silencer® select Product # s28935,s28936). Western blot analysis (Fig. a) was performed using whole cell extracts from the WNT4 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with WNT4 Antibody (9H2L10), Recombinant Rabbit Monoclonal (Product # 701857, 1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/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 WNT4.