This Blomia tropicalis BLOT2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of BLOT2 protein (Cat: 69062-A08H) from the overexpression lysate was verified.
A DNA sequence encoding the BLOT2 (AAQ73483.1) (Met1-Asp144) was expressed with a polyhistidine tag at the C-terminus.
The recombinant BLOT2 consists of 137 amino acids and predicts a molecular mass of 15 kDa.
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.