This Mouse PILRA overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PILRA protein (Cat: 50234-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse PILRA (NP_705730.1) extracellular domain (Met 1-Val 197) was fused with a polyhistidine tag at the C-terminus.
The recombinant mouse PILRA consists of 177 amino acids and has a predicted molecular mass of 20.2 kDa. As a result of glycosylation, the apparent molecular mass of rm PILRA is approximately 35-45 kDa in SDS-PAGE under reducing conditions.
Mouse PILRA 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.