In general ELISA requires to incubate at 37 degree up to 1 to 2 hrs for secondary HRP antibody to bind to antigen. For some special ELISA requires antigen overnight incubation at 37 degree, like in my case HMGB1-ELISA require to incubate at 37 degrees for 20 to 24 hrs. So, I would think in your case ELISA should be okay to incubate 2-3 hrs at 37 degree.

Hi,

Yes you could.

As in few steps of elisa we incubate our microtitre plate at 37 degree like when add serially diluted serum in solid phase binding assay of complement system. But before placing at 37 degree please take a look over the required temp for that specefic step, and if only room temp is recomended then first place it in an incubator having temp in between 20 to 23 degree & if still it doesn't work then try at 37 degree. Because high temp could denature your molecules.

If you still need any help or guidance you can inquire.

Thanks,

Mubasher

& one more imp point, the time you incubate is 90 mints (recomended in majority of the steps of solid phase binding assay) but still i suggest you to check suggested time & temp in protocol.

Thanks,

Mubasher