MSH3 Human Over-expression Lysate

Ship at ambient temperature. Upon receiving, store the sample at -20°C. Lysate samples can be reconstituted with SDS Sample Buffer. Avoid repeated freeze-thaw cycles after reconstitution. Lysate samples are stable for 12 months from date of receipt when stored at -20°C.

HEK293T cells in 10-cm dishes were transiently transfected with MegaTran Transfection Reagent and 5ug TrueORF cDNA plasmid. Transfected cells were cultured for 48 hours before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM 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°C before shipping.