I am trying to quantitate the amount of recombinant P450 in membrane preps using the reduced Fe vs Fe-CO method and am having difficulty getting ANY peaks on my spectrum scan between 400 and 500 nm. I am using a Shimadzu UV-1601 spectrophotometer that has a back reference cell and forward sample cell. In addition, I am using sodium bisulfite (sodium dithionite) as my reductant. I am pretty certain that the dithionite is relatively old and has been kept at room temp without dessication so I am not too sure of its reducing power anymore. If anyone has used this model of spectrophotometer to measure P450 content please let contact me. Any suggestions would be helpful. Thanks.
Hallo Kevin
Have a look at the following article - it is on Research Gate.
Swart, P., van der Merwe, K.J., Swart, A.C., Todres, P.C., Hofmeyr, J.-H.S., 1993b. Inhibition of cytochrome P-450 by some naturally occurring acetophenones and plant extracts from the shrub Salsola tuberculatiformis. Planta Medica 59, 139–143.
See the profile of Pieter Swart: https://www.researchgate.net/profile/Pieter_Swart2/publications/ for more publications.
You can also contact my colleague at [email protected] as he is an expert in the field of P450s.
Hope it helps.
Regards
Marina
Thank you very much for your response and guidance to the article. I will let you know if it is of any help and will contact your colleague if I still am having problems or concerns.
Best,
Kevin
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