Detection of cytochrome b5 from the house-fly, Musca domestica: comparison of immunological and spectrophotometric methods.
Spectrophotometric assay of microsomal cytochrome b5 in house-flies produces different results depending on whether sodium dithionite or NADH is used as the reducing agent and whether or not detergent is present. Microsomes assayed for cytochrome b5 with dithionite in the presence of detergent gave the highest values, followed by dithionite alone, NADH plus detergent, and then NADH alone. Isopropanol treatment of microsomes extracted cytochrome b5 free of spectrophotometrically interfering cytochrome P-450. Studies using immunoblotting and rocket immunoelectrophoresis with polyclonal antisera raised against the purified cytochrome b5 showed that isopropanol treatment quantitatively extracted cytochrome b5.
Wheelock
G D
GD
Department of Entomology, Cornell University, Ithaca, NY 14853-0999.
Scott
J G
JG
D000433:1-Propanol; D000818:Animals; D015786:Cytochromes b5 / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D004227:Dithionite; D006793:Houseflies / Q000737:chemistry*; D015151:Immunoblotting; D007122:Immunoelectrophoresis; D009243:NAD; D010084:Oxidation-Reduction; D013056:Spectrophotometry, Ultraviolet
D009243:NAD; D004227:Dithionite; D015786:Cytochromes b5; D000433:1-Propanol