This Mouse Resistin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Resistin protein (Cat: 50488-M01H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse RETN (Q99P87) (Ser 21-Ser 114) was fused with the Fc region of human IgG1 at the N-terminus.
The recombinant mouse RETN/Fc is a disulfide-linked homodimer. The reduced monomer comprises 354 amino acids and has a calculated molecular mass of 38.6 kDa. The apparent molecular mass of the monomer is approximately 40 kDa in SDS-PAGE under reducing conditions.
Mouse Resistin 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.
Resistin is an adipocytokine, which has been studied for its role in insulin resistance and recently in inflammation. The RETN and CAP1 polymorphisms and gene expression may be potential biomarkers for breast cancer risk. Resistin (RETN), recently found to be relevant to inflammation and inflammatory disorders.
Verma S, et al. (2003) Resistin promotes endothelial cell activation: further evidence of adipokine-endothelial interaction. Circulation. 108(6):736-40.
Reilly MP, et al. (2005) Resistin is an inflammatory marker of atherosclerosis in humans. Circulation. 111(7):932-9.
Steppan CM, et al. (2002) Resistin and obesity-associated insulin resistance. Trends Endocrinol Metab. 13(1):18-23.