This Mouse WIF1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of WIF1 protein (Cat: 50984-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse WIF1 (Q9WUA1) (Met1-Trp379) was expressed with a C-terminal polyhistidine tag.
The recombinant mouse WIF1 comprises 362 amino acids and has a predicted molecular mass of 39.8 kDa. The apparent molecular mass of the protein is approximately 44 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Mouse WIF1 HEK293 Overexpression Lysate: 使用指南
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
WIF1, also known as WIF-1 and wnt inhibitory factor 1, is a secreted protein that binds WNT proteins and inhibits their activities. It contains a WNT inhibitory factor (WIF) domain and 5 epidermal growth factor (EGF)-like domains. WNT proteins are extracellular signaling molecules involved in the control of embryonic development. WIF1 may be involved in mesoderm segmentation and can be detected in fish, amphibia and mammals. WIF-1 is a recurrent target in human salivary gland oncogenesis. Downregulation of WIF1 takes part in the development and progression of pleomorphic adenomas. WIF1 also is a tumor suppressor, and has been found to be epigenetically silenced in various cancers, specifically in nonfunctioning pituitary tumors. WIF1 are expected to have molecular function (protein tyrosine kinase activity) and to localize in various compartments (extracellular space, extracellular region).
WNT inhibitory factor 1
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Lin YC, et al. (2006) Wnt signaling activation and WIF-1 silencing in nasopharyngeal cancer cell lines. Biochem Biophys Res Commun. 341(2):635-40.
Queimado L, et al. (2007) WIF1, an inhibitor of the Wnt pathway, is rearranged in salivary gland tumors. Genes Chromosomes Cancer. 46(3):215-25.