Recombinant Anti-BDNF antibody [EPR1292] (ab108319)
In Abcams protocol and customer reviews it is stated that the incubation stage with this primary antibody should be around 16-18 hours at 4°C.
In our lab, some people have incubated for 1 hour at room temperature, with mixed results.
Is it critical to incubate for such a long period of time and at 4°C with primary antibodies in general? And does anyone have experience with this anti BDNF antibody specifically?
Many thanks.
I cannot comment on that antibody specifically but I have done hundreds of westerns with a variety of antibodies and I have always used a 1h incubation at room temperature, with gentle agitation.
Possible explanations for the long incubation time are (i) that Abcam when testing its Abs has a default overnight procedure to avoid wasting daytime hours on incubations, then 4oC temp also makes sense to prevent evaporation (if airtight containers are not being used). (ii) the antibody has a particularly slow on rate.
You will need to do a straight comparison with identical blots using the two incubation times if you want to know the answer.
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