Tyrosine hydroxylase immunostaining - (Jun/13/2007 )

Hi all,
Can someone give me a good working protocol for tyrosine hydroxylase immunostaining on mouse brain sections? - the dilution for primary antibody, the best method to do antigen retrieval etc etc

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We used this protocol for antigen retrivel.

Incubation in Tris-buffered saline (20 mm Tris,pH 9.0 and 136 mm NaCl) at 60C for 4-6hrs.

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Hi,
I have used citrate buffer for antigen retrieval - 1 hr at 65C, followed by endogenous peroxidase quenching for 1/2 hr and then blocking with BSA/milk powder. Then 72hrs primary (anti TH - 1:100) at 4C, followed by 2hr secondary. It still did not work. The primary that i used was from sigma. can someone suggest a better protocol?
My3

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