This Mouse CD229 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD229 protein (Cat: 50504-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse Ly9 (NP_032560.2) (Met 1-Arg 453) was expressed, with a polyhistidine tag at the C-terminus.
The recombinant mouse Ly9 consists of 417 amino acids and has a predicted molecular mass of 46.5 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rmLy9 is approximately 65 kDa due to glycosylation.
Mouse CD229 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.