[Histonet] Antibody diluent

[Rene J Buesa]

Hi Gyula:
To prepare the antibody diluent you will need also a preservative (use sodium azide). The formula is:
Bovine serum albumen ---- 1 mL
PBS with 0.05% Tween 20 ---99 mL
sodium azide --- 100 mg
René J.

--- On Thu, 6/4/09, Szabo, Gyula <Gyula.Szabo <@t> ucsfmedctr.org> wrote:


From: Szabo, Gyula <Gyula.Szabo <@t> ucsfmedctr.org>
Subject: [Histonet] Immunhistochemical practise
To: "'histonet <@t> pathology.swmed.edu'" <histonet <@t> pathology.swmed.edu>
Date: Thursday, June 4, 2009, 11:55 AM


Hi,

My name is Gyula Szabo from UCSF. I have a research position and I must use some immunhistochemical process.
I do it manual and I would like to make by myself almost every chemical solution.
I have some method but I need some correct exam because I could not find any information about it.

I need the correct method of making antibody diulent and protein blocking solution.

I know that the conponents are PBS buffer and 1% BSA but I do not how much is it need.

If I make for 1000 mL antibody diulent, how much is BSA or good idea if for 1000 mL PBS add 1 g BSA?

I have the fallowing method for protein blocking: 5 g non fat dry milk (like BSA) + 100 mL PBS buffer, then ulta centrifuga using at 1 hour than filtering.

If this method is correct how long time the blocking process and must I use for this step the humidity chamber?


I am looking forward to hearing from you.

Thank you.

Gyula


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