I have done a qualitative ELISA-analysis, and I need some help with my calculations. I tested my protein against different receptors. I used a plate reader for the absorbance (450nm) of the wells, and I also have the concentration (ng/μL) of the different receptor-solutions. I have fixed the absorbance values so they correlate with the time it took for the solutions to turn blue (HRP, TMB and HCl).
I wonder if anyone knows how I can combine these values and make it into one factor, so I can directly compare the affinity values between the different receptors and the controls.
The factors I have to play with are: Concentration (ng/μL), absorbance values, volume (L) and molar mass (g/mol) of the different receptors and controls.
I already used the factors molar mass, volume and concentration to calculate moles of the receptor-solutions. Maybe that's something to use.
Best,
Sigrid.
Hi Sigrid Berg . For a qualitative ELISA you can use EC50, which is 1/2 of the maximum binding (or saturation). This would be a good comparative if you are using the same antibody against the same protein.
good question ... the answer please refer to the article ... Miles, et al
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