I am confused whether to subtract the absorbance values of samples with the absorbance values of field blanks or do I subtract those after obtaining the concentration?
Substract the blank absorbance values, not the calculated concentration.
I must sadly and repeatedly say that the notorious ELISA and RIA methods are not quantitative. Notorious PCR method is also not quantitative. Quantitative determination methods should have a linear calibration line inserting to the zero point.
Instead, please use the quantitative HPLC-photometric method to determine the amount of molecules.
ELISA method gives c.a. 2,000-fold lower values than HPLC-photometric method (please compare the two files; Fucoidan ELISA and JCB Fucoidan transport).
Dear Ms./Mr. Asri,
It is common that if the sample OD at 405 nm was greater than the mean of the non infected controls plus 3 times their standard deviation, the sample is possibly infected with the pathogen of interest. However, sometimes the interpretation is not so easy and you will have to do PCR, IC- or RT-PCR according to your work. Bests, Masoud
Yes, subtract the absorbence values and not the concentration
Dear Mr./Ms. Asri,
You don't need to subtract. Elisa Reader normally does this if programmed properly......
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