Hello everyone.
I need to test a reaction mix against bacteria, to state its inhibitory potency: how can I stop the reaction? I tried filtration using MicroCons, but part of the product is lost, while heating doesn't seem to get rid of the enzyme.
Dear Samuele
is the enzime added in The cultire media ?
in this case what about some cicle of centrifugation and resuspention in fresh media with out enzime to wash it away?
ciao
Manuele
Dear Manuele,
Enzymatic reaction is carried out in phosphate buffer, then the mix is centrifugated, surantant is added to culture media and bacteria are inoculated. I was thinking to carry out the reaction directly in the the culture media, but I'm concerned about its efficacy.
Grazie!! Samuele
You may chill the reaction mixture to inhibit substrate enzyme reaction. Are u checking the intracellular or extracellular activity of bacterial cells? Must elaborate which enzyme you intend to assay.
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