||Over-expression cells lysed in modified RIPA buffer.
||HEK 293 cells
||1 vial of 100 µg gene specific transient over-expression cell lysate in RIPA buffer1 vial of 100 µg empty vector transfected control cell lysate in RIPA buffer1 vial of 250ul 2xSDS Sample Buffer (4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromphenol blue, 100mM DTT)
||Transfected cells were cultured for 48hrs 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 (Sigma), 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.
||Lysate samples are stable for 12 months from date of receipt when stored at -20°C. Avoid repeated freeze-thaw cycles.Lysate samples can be diluted with 2xSDS Sample Buffer provided. After dilution, the protein sample should be aliquoted and stored at -20°C for long term storage. Prior to SDS-PAGE fractionation, boil the lysate for 5 minutes.