This Mouse PLGF/PGF overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PLGF/PGF protein (Cat: 50125-MCCH) from the overexpression lysate was verified.
A DNA sequence encoding the mouse PGF (P49764) (Met1-Pro158) was fused with five amino acids (DDDDK) at the C-terminus was expressed and purified.
The recombinant mouse PGF consists of 146 amino acids and has a calculated molecular mass of 16.6 kDa. The recombinant protein migrates as an approximately 28 kDa band in SDS-PAGE under reducing conditions.
Mouse PLGF/PGF 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.