This Mouse Serpina1d overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Serpina1d protein (Cat: 51172-M02H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse Serpina1a (XP_003945711.1) (Met1-Lys413) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant mouse Serpina1a comprises 627 amino acids and has a predicted molecular mass of 70.2 kDa. The apparent molecular mass of the protein is approximately 67-77 kDa in SDS-PAGE under reducing conditions due to glycosylation.
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.