1 vial of 100 µg gene specific transient over-expression cell lysate in RIPA buffer 1 vial of 100 µg whole HEK293T cell lysate in RIPA buffer 1 vial of 250ul 2xSDS Sample Buffer (4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT)
Storage
The lysate is shipped with dry ice. Upon receiving, store the sample at -80°C. Also after dilution, the protein sample should be aliquoted and stored at -80°C for long term storage. Avoid repeated freeze-thaw cycles. Lysate samples can be diluted with 2xSDS Sample Buffer provided. Lysate samples are stable for 12 months from the date of receipt when stored at -80°C.
Shipping
Dry Ice
Preparation
HEK293T cells in 10-cm dishes were transiently transfected with MegaTran Transfection Reagent (TT200002) and 5ug TrueORF cDNA plasmid. 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 (Thermo Scientific Inc.).
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