. Author manuscript; available in PMC: 2017 Jul 1.

Published in final edited form as: Curr Protoc Neurosci. 2016 Jul 1;76:2.12.1–2.12.37. doi: 10.1002/cpns.1

Table 2.12.3.

Time Required for each Basic Protocol Before and After Primary Antibody Incubation

Basic Protocol Number	Before Primary Antibody^a	After Primary Antibody^b
1	1 hr 40 min	4 hr 20 min
2	1 hr 40 min	5 hr
3	1 hr 40 min	7 hr
4	1 hr 40 min	9 hr 5 min
5	18 min^c or 5 min^d	50 min
6	18 min^c or 5 min^d	58 min
7	18 min^c or 5 min^d	1 hr 43 min
8	18 min^c or 5 min^d	1 hr 23 min^e or 2 hr 16 min^f

All times are calculated without treatment for endogenous peroxidase. If sections need to be treated with with phenylhydrazine or hydrogen peroxide, add 25 minutes for Basic Protocol 1 or 9 minutes for Basic Protocol 5.

Times do not include substrate development, mounting sections onto slides, or dehydrating and coverslipping.

For tissues fixed in paraformaldehyde:acrolein.

For tissues fixed in paraformaldehyde alone.

For detection using NiDAB (or DAB) substrate

For detection using SA-fluorescence