Unpurified cell lysate. HEK293 cells transfected with expression vectors harboring target gene were harvested and washed with PBS twice. The cell pastes were re-suspended with ice-cold PBS containing mammalian cell protease inhibitor cocktail and further lysed with freeze-thaw cycles. After clarifying with 20,000 g centrifugation at 4°C for 30 min, the lysate was aliquoted, lyophilized, and stored at -80°C immediately. Protein concentration was determined by BCA kit (Thermo Scientific, Inc.) using BSA as protein standard. The gene overexpression in lysate was confirmed by Western blotting using anti-His tag antibody and/or target-specific antibodies and the lysate derived from HEK293 cells transfected with the empty expression vector was used as a negative control.
Molecular Weight (kDa):
Recombinant protein product has a calculated molecular mass of 14. The actual molecular weight may increase slightly due to the potential post-translational modifications (PTMs).
Lyophilized from a 0.2 um filtered solution in PBS (pH 7.4) containing mammalian cell protease inhibitor cocktail
Determined by BCA protein assay
Briefly spin the vial and bring the contents to the bottom prior to opening. It is recommended to reconstitute at 0.5 - 1 mg/mL with sterile deionized water.
Stability & Storage
Ice packs. Including one vial of HEK293 cell lysate transfected with empty expression vector.
Upon arrival, the lyophilized protein may be stored for 2 weeks at 4°C. For long term storage, it is recommended to store desiccated below -20 °C in a manual defrost freezer. Following reconstitution, the protein may be stored for 2 weeks under sterile conditions at -20 °C. For long term storage, it is recommended to make appropriate aliquots and store at -80 °C. Avoid repeated freeze-thaw cycles.