This Cynomolgus 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: 90108-C02H) from the overexpression lysate was verified.
A DNA sequence encoding the cynomolgus IFNA13 (G7NFW4) (Met1-Glu190) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant cynomolgus IFNA13 is a disulfide-linked homodimer. The reduced monomer comprises 407 amino acids and has a calculated molecular mass of 46.4 KDa.The apparent molecular mass of the protein is approximately 47 KDa respectively in SDS-PAGE.
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.