Human LIPF Baculovirus-Insect cells Overexpression Lysate: 产品信息
This Human LIPF overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of LIPF protein (Cat: 15652-H08B) from the overexpression lysate was verified.
A DNA sequence encoding the human LIPF (AAI12273.1) (Met1-Lys398) was expressed with a C-terminal polyhistidine tag.
The secreted recombinant human LIPF consists of 389 amino acids and predicts a molecular mass of 44.6 KDa. The apparent molecular mass of the protein is approximately 45 KDa in SDS-PAGE under reducing conditions.
Human LIPF Baculovirus-Insect cells 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.