This Human FAM19A4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of FAM19A4 protein (Cat: 11165-H01H) from the overexpression lysate was verified.
A DNA sequence encoding the human FAM19A4 (Q96LR4) (Ser36-Arg140) was expressed with the Fc region of human IgG1 at the N-terminus.
The recombinant human FAM19A4 /Fc comprises 365 amino acids and has a predicted molecular mass of 40.1 kDa. The apparent molecular mass of the protein is approximately 43 kDa in SDS-PAGE under reducing conditions.
Human FAM19A4 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.