Hello everyone,
I have developed a NCI-ELISA from scratch and what I want to do is to quantify a protein produced by a microorganism in different media. For this reason I am doing the Std curve in different media.
In the attached document you can see 2 std curves: they both have the same concentrations of pure peptide, same protocol, same plate. The only difference is that one is in CB and the other one is in CB+Media1.
They both look good to me, however, they are very different.
My questions are: why are they so different and what can I do about it?
Any comment or suggestion is very welcome
Thanks a million in advance
Hello
I saw your document. I drew standard curve by making serial dilution of known protein by certain concentration. It is not necessary to see the color of Media 1 in the wells, you should stop the ELISA before seeing non specific reaction in the negative control well and after seeing the color in the positive control. When you use Log of concentrations, the curve will be transformed to the line (whereas when you use the serial dilution, the curve will be sigmoid shape). So,you should add trendline by right click on the curve. Tick R2 and formula.
The greater R2 , the exacter and better standard curve.
Be successful
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