[Histonet] BSA in PBS

Rene J Buesa rjbuesa <@t> yahoo.com
Wed Mar 29 07:30:27 CST 2006


Hello Louise:
  Bovine albumen at 1% but if you are going to prepare a solution that will last you for more than 1 week it would be good to add 100 mg of sodium azide. Also the PBS should have 0.05% Tween 20
   
  The only thing that I could recommend is lieu of all the tings you mention is to have a large metal pot filled with water and a strong heat source (like a Bunsen burner). Place the container with the buffer (without the slides) and heat the water to boiling point. When both the water in the pot and the buffer in the container reach isothermal conditions (100ºC depending on atmospheric pressure) add the slides. Make sure that there is enough water in the pot to keep the water boiling through the duration of HIER.
  This (very old) arrangement for a water bath is known in some countries as "Mary (the alchemist) bath" and for sure will allow a good and isothermic HIER. It will require more time, though.
  Hope this will help you!
  René J.

louise renton <louise.renton <@t> gmail.com> wrote:
  Hello all,

would someone please remind me of the concentration of Bovine serum
albumin to put in PBS buffer to reduce background staining (IHC)?

Also, if I don't have a microwave, pressure cooker, rice/vegetable
steamer, or autoclave - whats another way to do heat mediated antigen
retrieval?

Best regards and thanks in advance

--
Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
"Does a conceited cowboy, only ride on pompous grass?.

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