I am doing my master’s thesis about cytokines and I’m using the R&D systems DuoSet ELISA kit. My problem is that I don’t know what kind of standard curve is should be using. The plate reader (Fluostar omega) gives linear fit, but I think it does not work in this case because lower absorbance standard points are turning negative, which have an effect on concentrations.
What kind of standard curves are you using for ELISA or what kind of standard curve are you recommending to use? Maybe Hyperbola fit?
Thanks in advance for your answers!
I have used R&D ELISA Kits before and have always used a linear fit for my standard.
If your samples have high cytokine concentrations, I would suggest that you dilute your samples so that they land in a concentration range where your standard curve is the most accurate. You can also adjust the concentrations of your standard samples to more accurately represent the concentration range your samples are in.
Good luck!
In general, you may use any curve, which fits your data properly. Since ELISA curves vary with experimental conditions, there is no curve for all cases. Take care that your sample signals are in the range you calibrated, otherwise a dilution of the samples might be necessary, as mentioned by Jan Philipp Dobert
Dear I suggest you follow the protocol provided by the manufacturer of the ELISA kit.
Hi Ira! In our work we use the program CurveExpert, which itself offers the optimal curve.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA