Hi all,
I've been reading and looking for some protocols of storage of erythrocytes for enzymatic antioxidant (SOD,CAT, GPx) measurement, however, I found plenty of information that bring me doubt. My questions are:
1) After washed the erythrocyte, should I storage it at -80 with some buffer ?
2) How long can I storage ( one year or months)?
Would you have some protocol to share with me this method?
Many thanks,
Whenever you try to keep enzymes in their active conformation, the storage temperature should be -80. I once checked, how enzyme activity decreases at -20 C freezer, and in 1 month activity decreased 20-50 %.
Separate plasma by centrifugation of whole blood at 1500 X g removed upper layer plasma take erythrocytes fraction stored at _ 80 C up to 45 days as separate aliquot so each time use one aliquot thawed and assay for the dsired enzymes
good luck
Wash erythrocytes with osmolar saline thrice and prepare hemolysate .You can store at -20 for 1 week or 2-8 for 3 days .Don't keep it fir more than 1 week for enzymatic analysis
1- he storage temperature should be -80 .
2- as long u storage as the activity of enzyme will decrease
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