In sandwich ELISA, R&D kit insert is mentioned 20 min is the incubation time for Streptavidin-HRP conjugate.
If we increase the Incubation time from 20 min to 120 min what all the effects we can expect on OD?
This cannot be predicted from your information. In many cases, you get a higher OD after longer incubation time. In the case of the streptavidin-HRP conjugate, a small improvement in assay sensitivity (this is what you mean with a potentially higher OD?) may be observed. However, this has to be verified experimentally.
I agree with Dr. Weller.
What you should also keep in mind is that a way longer incubation time with Strep-HRP usually results in a higher background and your blanks will give too high signals, which eventually not really ups your sensitivity.
If you aim for a higher sensitivity you should rather increase the incubation time of your target protein and/or your primary biotin antibody.
In my experience slightly increasing all the incubation times results in higher sensitivity and good ODs, rather than just extremely increasing only one.
Good luck!
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA