This Mouse IFN-alpha 13 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IFN-alpha 13 protein (Cat: 50670-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse IFNA13 (Q80SU4) (Met1-Glu189) was expressed with a C-terminal polyhistidine tag.
The recombinant mouse IFNA13 comprises 177 amino acids and has a predicted molecular mass of 20.5 kDa. The apparent molecular mass of the protein is approximately 26-31 kDa in SDS-PAGE under reducing conditions.
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.