Hi,
I was wondering how could I validate whether a solution with 0.5 mM EDTA; pH 9.0 & 10 mM Tris-Cl may have any absorbance values at 492 nm ( i rather mean if I can check it online)
Additionally do any of these products have any known interaction with HRP and 0-phenylene diamine reaction?
I am doing ELISAs with DNA as epitope. therefore I isolate DNA from some cells and then load these on the plate. Anyhow I have noticed that the lower the amount of DNA in my preps, in other words, the more volume I take from a given prep I get unexplainable higher values on my ELISA reads for that prep. As said this values cannot be explained by anything, but strongly correlate with this particular volume issue. Thus I am wondering whether if it is possible that actually my residual elution buffer from the DNA preps is somehow messing with the reads.
Thanks a lot!
Hi Inigo,
The compounds you mentioned are not UV absorbing, but sometimes they do if they contain some impurities. I recommend to measure this solution yourself to check if it absorbs UV or not (as a blank).
Good luck!
Hi Fatma,
Thanks for your reply! anyways I was not asking for UV absorption. the DNA prep measurements doent bother me because i have the buffer as blank and I actually know that our nanodrop is very good with a 0.4 low detection limit, which is very good. its more if somehow later on during elisa this elution buffer carryover could somehow mess with the reaction or the reads or something.. I know that the prptocol has lots of washing stepts and you would not expect anything to "survive" so long in the plate so it actually is a silly hipothesis, but i do not have a better one..
Maybe I can try to elute with water simply and make life easier
Any thought are welcome!
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