Anti-Wilms′ Tumor Antibody, NT clone 6F-H2, Ascites Free

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.

Immunohistochemistry Analysis: An 1:250 dilution from a representative lot detected Wilms tumor protein in human kidney tissue.

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).

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.

Evaluated by Western Blotting in Jurkat cell lysate.

Western Blotting Analysis: 1.0 µg/mL of this antibody detected Wilms tumor protein in 10 µg of Jurkat cell lysate.

~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.

Format: Purified

Concentration: Please refer to lot specific datasheet.