Help!!!

[Marianapb]

I am with an Elisa for detect activin A, we should let it overnight with the capture antibody at room temperature but i let it at 4°C, should I wait 12 hours letting it at room temperature or what shoud i do? somebody told me that I could put it at 37°C 1 hour, that wont affect the assay?
Please help!!!

[Elisadeveloper]

It will probably benefit from some time at RT, at worst you ODs might be a little lower than otherwise. Which kit are you using?

[Zagami]

Isn’t important that you possess a lot ideas, but that you live one of it. Each researcher must titrate a particular antigen to obtain standardized coating conditions. If your antibody anti-activin A is polyclonal or monoclonal (rh-FS228, 2F8:6H5, etc…) or also from chicken derived (IgY), is necessary that different operative conditions must be tried, for to obtain a detection limit, with which 95 % of the normal and pathological values of the samples are correctly identified. For Mab afore mentioned I wish to acquaint the coating optimized conditions :
Mab concentration : 4-10 µg/ml
Coating buffer : sodium bicarbonate 0.2 M at pH 9.4
Incubation time : 16 h (overnight)
Incubation temperature : 25 °C (room)
Shaking : optional, gently at 500-600 rpm

Good luck and to be on hand