10 minutes; take 200 µL of supernatant, add an equal volume of alkaline extract ; Mix well, centrifuge
at 10,000g at 4°C for 10 min, take the supernatant, and store on ice for testing.
Extract NADPH: Take 0.1 mL of serum (slurry), add 0.5 mL of alkaline extract, boil for 5 minutes
(cover tightly to prevent water loss), after cooling in an ice bath, centrifuge at 10,000g at 4°C for 10
minutes, take 200 µL of supernatant, and add an equal volume of acidic extract ; Mix well, centrifuge
at 10000g at 4°C for 10min, take the supernatant and store on ice for testing.
2. Tissue
Extract NADP+: Weigh about 0.1g of tissue, add 0.5mL of acidic extract, grind in ice bath, boil for
5min (cover tightly to prevent water loss), after cooling in ice bath, centrifuge at 10,000g at 4°C for
10min, take 200µL of supernatant, add an equal volume of alkaline extract ; Mix wel, centrifuged at
10,000 g at 4°C for 10 min, and the supernatant was taken and stored on ice for testing.
Extract NADPH: Weigh about 0.1 g of tissue, add 0.5 mL of alkaline extraction solution, grind in an
ice bath, boil for 5 min (cover tightly to prevent water loss), after cooling in an ice bath, centrifuge at
10,000 g at 4°C for 10 min, take 200 µL of supernatant, and add an equal volume of acidic extract ;
Mix well, centrifuged at 10,000 g at 4°C for 10 min, and the supernatant was taken and stored on ice
for testing.
3. Bacteria or cells
Extract NADP+: Collect 5 million cells or bacteria, add 0.5mL of acidic extract, ultrasonically disrupt
for 1min (intensity 20% or 200W, ultrasonic for 2s, stop for 1s), boil for 5min (cover tightly to prevent
water loss), cool in ice bath, 10000g centrifuge at 4°C for 10min, take 200uL of the supernatant into
another new centrifuge tube, add an equal volume of alkaline extract to neutralize, mix well,
centrifuge at 10,000g at 4°C for 10min, take the supernatant and store it on ice for testing .
Extract NADPH: Collect 5 million cells or bacteria, add 0.5mL alkaline extract, ultrasonically
disrupt for 1min (intensity 20% or 200W, ultrasonic for 2s, stop for 1s), boil for 5min (cover tightly to
prevent water loss), cool in an ice bath, Centrifuge at 10,000g at 4°C for 10min, take 200uL of the
supernatant into another new centrifuge tube, add an equal volume of acidic extract to neutralize, mix
well, centrifuge at 10,000g at 4°C for 10min, take the supernatant, and store it on ice for testing .
Determination