OD > 0.300
Standard Deviation> 0.08
Negative control has high OD.
Any advice I will listen!
I dont understand properly how is your ELISA performed. the results are also unclear for me. could you add more details?
Yes also if you are using commercially available mAbs we may be familiar with?
If it helps anyone who is wondering for a plate ELISA, regardless of type should be tested with ranges of anti-p24 from 1-10ug/mL
You want to develop a double antibody sandwich for detection of p24. First you need to make sure that you have a maximal surface concentration of capture antibody (monolayer), second the optimal conjugate concentration (complete reaction of captured Ag). This is done by chess board titration. (see textbooks) The detection limit of such optimised ELISA is (provided the nonspecific binding is under control) limited by two facts:
1. binding constants of the antibodies -> take more affine antibodies, try other adsorption methods (immobilisation can reduce binding constant)
2. generated signal per binding event-> try other labels-> the best I know is poly HRP (increases the signal 10-100 times)
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