Western blot analysis of extracts from SK-OV-3, OVCAR3, and K-562 cells using WT1 (D6M6S) Rabbit mAb.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
WT1 (D6M6S) Rabbit mAb recognizes endogenous levels of total WT1 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val306 of human WT1 protein.
Wilms' Tumor 1 (WT1) is a transcription factor named from Wilms' Tumor 1, an embryonal malignancy of the kidneys that is caused by mutations in the WT1 gene (1). It is highly important in development, particularly of the genitourinary system, and mutations and dysregulation of expression of WT1 result in a variety of syndromes affecting the genitourinal system and other tissues (2-5).
WT1 has a myriad of biological functions and a host of interacting partners and target genes (6). It can behave as a transcriptional activator, or a repressor, and can act as an oncogene or a tumor suppressor (7). It exerts influence over the epigenetic landscape, and also has post translational influence of gene expression through RNA interactions (8). The diverse biological roles of WT1 have been attributed to the existence of multiple isoforms and post translation modifications of the protein (9).
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