This Human NPTXR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NPTXR protein (Cat: 16070-H07H) from the overexpression lysate was verified.
A DNA sequence encoding the human NPTXR (NP_055108.2) (Ala24-Ala500) was expressed with a polyhistidine tag at the N-terminus.
The recombinant human NPTXR consists of 496 amino acids and predicts a molecular mass of 52.8 kDa.
Human NPTXR 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.