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Immunocytochemistry - Primary Ab incubation time

ICC primary antibody incubation

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#1 sc_queen

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Posted 21 January 2012 - 10:38 AM

Hello,

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?

Thanks!

Edited by sc_queen, 21 January 2012 - 11:01 AM.


#2 BioMiha

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Posted 22 January 2012 - 12:54 AM

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.

#3 sc_queen

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Posted 22 January 2012 - 01:23 AM

Thanks BioMiha! It helps.

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





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