This Mouse DKK3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DKK3 protein (Cat: 50247-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Ile 349) of mouse DKK3 (NP_056629.1) precursor was expressed with a C-terminal polyhistidine tag.
The secreted recombinant mouse DKK3 consists of 338 amino acids and has a calculated molecular mass of 38 kDa. As a result of glycosylation, the recombinant protein migrates as an approximately 65 kDa protein in SDS-PAGE under reducing conditions.
Mouse DKK3 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.
DKK3 (dickkopf related protein 3) is a member of the dickkopf-related family consisting of DKK1, DKK2, DKK3 and DKK4. It is a secreted protein, and also known as REIC (Reduced Expansion in Immortalized Cells). The DKK3 protein is proposed to function as a secreted tumor suppressor since it is downregulated in a number of cancer cells and prostate cancer tissue and may be a promising candidate molecule for therapeutic interference. DKK3 protein is also a negative regulator of beta-catenin and its downregulation contribute to an activation of the beta-catenin signaling pathway.
dickkopf WNT signaling pathway inhibitor 3
Canonical Wnt Pathway
Yue W, et al., 2008. Downregulation of Dkk3 activates beta-catenin/TCF-4 signaling in lung cancer. Carcinogenesis, 29 (1): 84-92.
Ding Z, et al., 2009. Promoter methylation and mRNA expression of DKK3 and WIF-1 in hepatocellular carcinoma. World J Gastroenterol, 15 (21):2595-601.