IHC of the brain secions

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Amitjais
Amitjais's picture
IHC of the brain secions

Hi Pudir,
M fine. What abt u?
I need a standard  protocol for the HRP staining of TH in the brain sections. Do the thickness of the section i.e. 20 micron needs some more time of permeabilization of staining? and how to get reduced background and specific staining. Kindly send me the protocol.

Arvind Singh Pundir
Arvind Singh Pundir's picture
 Hi Amit,

 Hi Amit,
thickness do matter at times for certain antigens i am using 50 micron sections with 0.3% Trit X-100 for 20 minutes in Quenching step as well as in primary antibody preparation(incubation) step to reduce the background staining that may be due to peroxidase , use 2 % H2O2 for 20 min at RT i have used TH from chemicon at the dilution of 1:250 overnight at  4 degree celcius rest the protocol is same as usual one for frozen sections if you are using the same
link for a general protocol used for frozen sections :
http://www.scientistsolutions.com/a115-protocol-standard+immunohistochemical+staining+procedure+for+frozen+sections.aspx