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Radioactive thiols in the eluate were identified by comparison with the fluorescence peaks of mBBr derivatives and by S-cysteine, S-glutathione and S-sulphate standards.
After removal of the methanol of the combined eluates under reduced pressure, the pH of the aqueous fraction was adjusted to 3.0 and partitioned three times against ethyl acetate at pH 3.
Aliquots of eluates containing proteins were rapidly frozen in liquid nitrogen and stored at - 80°C for protein determination and enzyme activities.
This elution step was repeated twice and eluates were pooled.
Concentrated CKs were eluted from the cartridges with ethanol and the eluates were evaporated to dryness.
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