The WNT gene family consists of structurally related genes which encode secreted signaling proteins. These proteins have been implicated in oncogenesis and in several developmental processes, including regulation of cell fate and patterning during embryogenesis. WNT4 is a member of the WNT gene family, and is the first signaling molecule shown to influence the sex-determination cascade. It encodes a protein which shows 98% amino acid identity to the Wnt4 protein of mouse and rat. This gene and DAX1, a gene known to antagonize the testis-determining factor, play a concerted role in both the control of female development and the prevention of testes formation. WNT4 and another two family members, WNT2 and WNT7B, may be associated with abnormal proliferation in breast tissue.
Domain Information: Wnt family of developmental regulators
Gene Ontology Terms: cell-cell signaling; development; extracellular matrix structural constituent; frizzled-2 signaling pathway; signal transducer activity
Application(s)
Peptide ELISA: antibody detection limit dilution 1:32,000. Western Blot: Approx 35kD band observed in lysates of Human Testes and Ovary (calculated MW of 39.1kD according to NP_110388.2). Recommended concentration: 0.03-0.1ug/ml.
Staining (0.03ug/ml) of a) Human Ovary and b) Human Testes (RIPA buffer, 35ug total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence. Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Peptide Blocking
Corresponding peptide is available for Peptide Blocking studies. See W3021-01-P. Antibody is typically 0.5mg/ml and peptide is supplied as a 100ul pellet. When peptide is reconstituted in 200ul water, the concentration would also be 0.5mg/ml. To start, the best ratio would be 1:1 (molar excess of peptide relative to antibody when identical volumes are mixed). Mix equal volumes of peptide and antibody at the required dilution and leave at ambient temperature. It is best is to have two identical blots to be incubated with equal amount of antibody, but one with the antibody pre-adsorbed to the peptide for 20min. Then incubate and develop the two blots in parallel.
Immunogen
C-SNWLYLAKLSSVGS, internal region
Form
Supplied as a liquid in Tris saline, pH 7.2, 0.5% BSA, 0.02% sodium azide.
Purity
Purified by immunoaffinity chromatography.
Specificity
Recognizes Human, Mouse, Rat Wnt4