[Histonet] Immunhistochemical practise

[Szabo, Gyula]

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