Protocol Online logo
Top : New Forum Archives (2009-): : Immunology

Immunocytochemistry - Primary Ab incubation time - (Jan/21/2012 )


I am new to ICC and have a basic question. How do you determine incubation time and temperature for antibodies? It seems to vary a lot, esp. for primary, ranging from 1 hr at 37C to 1-2 days at 4C...What works better? Or does it depend on which proteins you are staining for?



I suppose the signal as a function of time is essentially much like the signal as a function of dose. It depends on the intrinsic properties of the Ab and you can improve the signal if you extend the time but there is a plateau. Incubating at 4 deg C is done to avoid losing the protein you are staining.


Thanks BioMiha! It helps.

Btw, you write very concisely yet clearly - something I really need to improve on!