This Mouse FCGR4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of FCGR4 protein (Cat: 50036-M08H) from the overexpression lysate was verified.
The amino acid sequence corronding to Met 1-Gln 203 of mouse FCGR4 (NP_653142.2) was fused with a polyhistidine tag at the C-terminus.
The recombinant mouse FCGR4 consists of 194 amino acids and has a predicted molecular mass of 22.4 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rmFCGR4 is approximately 25-35 kDa due to glycosylation.
Mouse FCGR4 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.