i have 102 samples and i have to estimate protein, gsh, lipid peroxidation, gst, gshpx, catalase and sod..... it will take weeks to finish all these assays, so, after making cell homogenates in pbs is there any way that i can store them without damaging the enzymes.... I am storing them at -80C and if i repeatedly freeze and thaw them is there chances that the enzymes might be damaged? how do i encounter this problem...
lysing in the presence of protease inhibitors, aliquoting, snap-freezing and then storing at -80 will be the best option. Adding additional components (eg. Glycerol) could stabilize the proteins from freeze-thawing damage, but you should check compatibility with your assay reactions. I am also not sure whether all the assays which you mentioned have the same tolerance levels for fresh versus frozen lysates (ex. Most of the GSH assays recommend storing pellets) !! Good Luck.
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