A DNA sequence encoding the human TNFRSF10A (NP_003835.2) extracellular domain (Met 1-Asn 239) was expressed, fused with a polyhistidine tag at the C-terminus.

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