This CyCMV CyCMV Glycoprotein B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CyCMV Glycoprotein B protein (Cat: 40369-V02H) from the overexpression lysate was verified.
A DNA sequence encoding the macaca fascicularis cytomegalovirus (CyCMV)(strain OT-1) glycoprotein B(gB) (ADV35145.1) (Met1-Asp680, with cleavage site mutated ‘RHKR’ to ‘TTQT’) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant macaca fascicularis cytomegalovirus (CyCMV)(strain OT-1) glycoprotein B(gB) consists 899 amino acids and predicts a molecular mass of 102.5 kDa.
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.