This Human MCEMP1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of MCEMP1 protein (Cat: 15757-H04H) from the overexpression lysate was verified.
A DNA sequence encoding the human MCEMP1 (NP_777578.2) (Lys107-Gln187) was expressed with the Fc region of mouse IgG1 at the N-terminus.
The recombinant human MCEMP1/mFc comprises 317 amino acids and has a predicted molecular mass of 36 kDa. The apparent molecular mass of the protein is approximately 43.7 kDa in SDS-PAGE under reducing conditions.
Human MCEMP1 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.