Compared to wild-type BoNT/X protease, both X(PC2)b, an earlier evolutionary intermediate with activity on both VAMP1 and procaspase-1, and X(PC) demonstrated more stringent substrate requirements (Fig
Briefly, sodium bicarbonate, glucose, sodium pyruvate, and all 20 amino acids (except cystine and serine), biotin (B4639, Sigma), and vitamin B12 (V6629, Sigma) were added to the DMEM medium (#D9800-28, USBiological, Salem, MA) to match standard IMDM, supplement with 15% human AB serum and 15% dialyzed FBS (#SH300790340, Cytiva Hyclone), along with 10 ng/mL interleukin-3, 50 ng/mL stem cell factor and 2IU/mL erythropoietin
But, its important to learn how from a healthcare provider
Vitamin B12 injections do away with all of that
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