This Mouse IL28A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IL28A protein (Cat: 51305-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse IL28A (NP_001019844.2) (Met1-Val193) was expressed with a C-terminal polyhistidine tag.
The recombinant mouse IL28A comprises 185 amino acids and has a predicted molecular mass of 21.1 kDa. The apparent molecular mass of the protein is approximately 28.5 and 24.7 kDa in SDS-PAGE under reducing conditions.
Mouse IL28A 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.