I am currently working on snake venom enzymes inhibition by crude plant extracts. I found one of the plant extracts working very effectively in different enzyme inhibitions. When I communicate the same results in reputed Journal, Reviewers asked for standards or positive controls. During the experiment, I have used venom activity in absence of plant extract as a control and also the proper blanks were maintained. Now my doubt is that, what is the necessity to have positive controls when I can easily say whether the experimental design is correct or wrong with controls itself? Could anyone explain this?
You are working working on possible snake venom enzyme inhibition by crude plant extracts. Is this the case? Your objective is to explore whether your extract contains inhibitors of these enzymes. What do you mean by "venom activity"? Are you using standard, purified commercial sources of snake venom enzymes such as phospholipase-A2 and phosphodiesterase or others?
Dear Sir,
Thank you very much for your queries on this problem. As you mentioned exactly, my objective is to check whether the plant extract could inhibit venom enzymes. I am currently working with crude venom. I havent purified any of venom enzymes. Venom activity means the activity of venom enzymes in vitro as well as en vivo in mice models. I am screening the plant extract against protease, phospholipase and hyaluronidase enzymes activity of crude venom.
Kindly give your suggestions to shape up my work more scientific.
Thanking you
Yours Sincerely
Gopi
< I am currently working with crude venom. I havent purified any of venom enzymes>.
I presumed that but wished to ask you in order to ascertain.
There is no need to purify crude venom enzymes; they are readily available commercially. Venom enzymes have been known for a long time, and phosphodiesterases have been utilized to hydrolyze nucleotides. Representative purified enzymes are commercially available, and these are the ones to be used for direct inhibition studies. They are relatively inexpensive.
I meant to ask you why you were using crude snake venom. Is it the cost factor of the commercial enzyme? Venom is a merry mixture of enzymes. In enzymology, to detect the potential inhibitory action of an agent, its effect on a reasonably purified, standard, enzyme is checked. Some factors in your crude venom might also have an inhibitory effect, in addition to that of your plant extract; and, in such a case you are not measuring the inhibitory potency. One would like to see whether you used a standard enzyme. Check the relevant volume of "Methods in Enzymology" for assaying one of these enzymes using a standard commercial enzyme reference); follow the procedure, and then add your plant extract and see the outcome.
On another issue, are safeguards in place at VIT for the usage and safe handling of potentially harmful and poisonous substances (biohazards) such as crude venom?
Wishing the best...
Inhibitors which covalently modify enzyme such as nitrogen mustards, aldehydes, haloalkanes, alkenes, Michael acceptors, phenyl sulfonates, or fluorophosphonates are potent inhibitors of enzymes..
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