General description
Wilms tumor protein (UniProt P19544; also known as WT33) is encoded by the WT1 (also known as DDS, FS, MEACHS, MESOM, NPHS4, WAGR) gene (Gene ID 7490) in human. The Wilms’ tumor gene WT1 was originally identified in the childhood kidney cancer Wilms’ tumor. The N-terminal region of WT1 protein contains a proline-rich region (a.a. 27-83) involved in transcriptional regulation, self-association, and RNA recognition, while its C-terminal region contains four zinc fingers (a.a 323-347, 353-377, 383-405, 414-438) that mediate DNA and RNA binding. The zinc finger domain of WT1 can bind to GC-rich sequences, such as the EGR-1 consensus sequence (5’-GCG(T/G)GGGCG-3’), the WTE motif (5′-GCGTGGGAGT-3′), or (TCC)n motif. Many genes responsible for cell growth and apoptosis, such as Bcl-2, Bcl-xL, BFL1, and c-myc, have been identified as downstream targets of WT1. There are four major alternatively spliced WT1 isoforms resulting from splicing at either or both of exon 5 (17AA) and exon 9 (KTS). All four major WT1 isoforms are overexpressed in leukemia and solid tumors and play oncogenic roles such as inhibition of apoptosis, and promotion of cell proliferation, migration and invasion.
Specificity
Expected to react with all spliced isoforms other than isoforms 6 and 9 reported by UniProt (P19544). Reactivity toward isoforms 6 and 9 has not been determined.
Immunogen
Epitope: Amino acids 1-173.
Recombinantt human WT-1 N-terminal fragment (a.a. 1-173).
Application
Immunoprecipitation Analysis: A representative lot co-immunoprecipitated CRE-binding protein/CBP together with Wilms tumor protein WT1 from the lysate of a T-SV40 immortalized human glomerular epithelial cell (HGEC) line (Drossopoulou, G.I., et al. (2009). Am. J. Physiol. Renal Physiol. 297(3):F594-F603).
Western Blotting Analysis: A representative lot detected Wilms tumor protein WT1 in the CRE-binding protein/CBP immunoprecipitate obtained from the lysate of a T-SV40 immortalized human glomerular epithelial cell (HGEC) line (Drossopoulou, G.I., et al. (2009). Am. J. Physiol. Renal Physiol. 297(3):F594-F603).
Western Blotting Analysis: A representative lot detected Wilms tumor protein WT1 in lysates from mouse E15.5 embryonic kidney and human melanoma cell lines A375, SK-MEL-28, and WM-266-4 (Wagner, N., et al. (2008). Pflugers Arch. Eur. J. Physiol. 455(5):839-847).
Immunocytochemistry Analysis: A representative lot immunostained the nucleus of methanol-fixed human melanoma A375 cells by fluorescent immunocytochemistry (Wagner, N., et al. (2008). Pflugers Arch. Eur. J. Physiol. 455(5):839-847).
Immunofluorescence Analysis: A representative lot immunostained the PCNA-positive nuclei of proliferating cells in formalin-fixed, paraffin-embedded human melanoma tissue sections by fluorescent immunohistochemistry (Wagner, N., et al. (2008). Pflugers Arch. Eur. J. Physiol. 455(5):839-847).
Immunohistochemistry Analysis: A representative lot immunostained glomeruli in formalin-fixed, paraffin-embedded normal human kidney and Wilms′ tumor sections (Wagner, N., et al. (2008). Pediatr. Nephrol. 23(9):1445-1453).
Immunohistochemistry Analysis: A representative lot detected vascular WT1 expression in 95% of 113 paraffin-embedded tumour tissues of various types. In most cases, nuclear WT1 staining of endothelial cells was seen (Wagner, N., et al. (2008). Oncogene. 27(26):3662-3672).
Immunohistochemistry Analysis: A representative lot immunostained the nucleus of perifollicular fibroblasts at the hair follicle in formalin-fixed, paraffin-embedded normal human skin sections. Most common melanocytic nevi do not express WT1, whereas Spitz nevi and dysplastic nevi show cytoplasmic WT1 staining. (Wagner, N., et al. (2008). Pflugers Arch. Eur. J. Physiol. 455(5):839-847).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
This Anti-Wilms′ Tumor Antibody, NT clone 6F-H2, Ascites Free is validated for use in Immunocytochemistry, Immunoprecipitation, Immunofluorescence, Immunohistochemistry (Paraffin), and Western Blotting for the detection of Wilms′ tumor protein.
Target description
~52 kDa observed. 49.19 kDa (isoform 1), 47.20 kDa (isoform 2), 47.51 kDa (isoform 3), 48.87 kDa (isoform 4), 34.45 kDa (isoform 5), 56.88 kDa (isoform 6), 55.21 kDa (isoform 7), 33.09 kDa (isoform 8) calculated. Uncharacterized band(s) may appear in some lysates.
Physical form
Format: Purified
Protein A purified.
Purified mouse IgG1κ in 0.02 M phosphate buffer (pH 7.6), 0.25 M NaCl and 0.1% sodium azide.
Storage and Stability
Stable for 1 year at 2-8°C from date of receipt.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.