Description
Transient overexpression lysate of ubiquitin specific peptidase 40 (USP40) with a C-Myc/DDK tag was expressed in HEK293T. HEK293T cells in 10-cm dishes were transiently transfected with cDNA plasmid. Transfected cells were cultured for 48 hrs before collection. The cells were lysed in modified RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1 mM EDTA, 1 x Proteinase inhibitor cocktail mix, 1 mM PMSF and 1mM Na3VO4), and then centrifuged to clarify the lysate. Protein concentration was measured by BCA kit. Cell lysates were aliquoted and stored at -20 centigrade before shipping. Components: 1 vial of 100 µg gene specific transient over-expression cell lysate in RIPA buffer 1 vial of 100 µg empty vector transfected control cell lysate in RIPA buffer 1 vial of 250 µL 2 x SDS Sample Buffer (4% SDS, 125 mM Tris-HCl pH 6.8, 10% Glycerol, 0.002% Bromphenol blue, 100 mM DTT)