Relative activity of NADPH oxidase activity bands (103–106 and 75–80 kD) in the fractionated supernatant and membrane proteins in the SDS in-gel assay
Fraction | Molecular | Soluble Protein | Relative Activity | Total Relative Activity (Soluble Protein × Relative Activity) |
---|---|---|---|---|
kD | μ fraction−1 | |||
Supernatant (203,000 g) | 103–106 | 813,000 | 0 | 0 |
77–80 | 0 | 0 | ||
Pellet (203,000 g) | 103–106 | 8,950 | 4 | 35,800 |
77–80 | 4 | 35,800 | ||
Lower phase | 103–106 | 350 | 1 | 350 |
77–80 | 3 | 1,050 | ||
Upper phase | 103–106 | 370 | 100 | 37,000 |
77–80 | 15 | 5,550 |
F2-a Extracts from 150 g of tomato leaves were centrifuged at 10,000g. The supernatant was subjected to centrifugation at 203,000g to yield the microsomal fraction. Lower and upper phases were partitioned from the microsomal fraction by the two-phase system and fractionated by SDS-PAGE as described in “Materials and Methods.” The activity gels were scanned and the results presented in arbitrary relative units.