Rapid and large-scale isolation of microsomal fraction of mouse liver by lyophilization and low speed centrifugation.
We could prepare the microsomal fraction of mouse liver, without using an ultracentrifuge but with a low speed centrifuge. The procedure includes 1) lyophilization of post-mitochondrial fraction (9,000 x g supernatant) of mouse liver, 2) powdering of the lyophilized sample, 3) the addition of 1.15 per cent potassium chloride solution or distilled water, which afforded microsomal aggregates, 4) sedimentation of microsomal fraction by low-speed centrifugation (20,000 x g, 20 min). The sedimented microsomal fraction showed normal contents of cytochrome P-450 and cytochrome b5, and gave a normal pattern on SDS polyacrylamide gel electrophoresis and normal electron microscopic feature. This method should be convenient for rapid and large-scale preparation of microsomes, especially for the preparation of cytochrome b5 and cytochrome P-450.
Iwata
N
N
Department of Forensic Medicine, Tokyo Medical College.
Mukai
T
T
Hara
S
S
Endo
T
T
Tomoda
A
A
D000818:Animals; D002498:Centrifugation / Q000379:methods*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D015786:Cytochromes b5 / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005612:Freeze Drying; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008862:Microsomes, Liver / Q000378:metabolism; D013997:Time Factors
D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; D007770:L-Lactate Dehydrogenase