Hello, I wish to know Whether it is good to carry out centrifugation in normal centrifuges instead of cooling centrifuges while doing enzymatic assays especially in cell culture.
it depends upon the need for a certian temperature to work in
so depends upon the protocol of the experiment and the cataloques of the reagents
Low temperature in cooling centrifuge first of all prevents the denaturation of your enzyme. If you planning to spin your protein sample at high g values and during prolonged time it may lead to significant increase of your centrifuge rotor temperature. Second, the low temperature slows down rate of reaction catalysed by your enzyme. Therefore, if you perform the centrifugation without cooling and the sample contains substrates for the enzyme after the process the substrate may be completely digested. This can be an issue if you planned to perform further enzyme kinetic studies on this sample.
In general, if you are dealing with extracts of human or other mammalian cells, it is likely that cooling will be important in order to slow down proteolytic degradation.. However, the best procedure is to assume nothing and actually test the requirements of your own system. If you can show that the protein you are working with is stable at room temperature over the relevant time period then you may be OK. There is also the possibility (and this is a very strong reason for CHECKING and not just assuming that you have to do what everyone else does) that you might have an enzyme that is COLD-LABILE - stable at 37C or maybe at room temperature but not at 4C. It happens! So try out the different conditions for yourself.
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