----- Original Message -----
Sent: Wednesday, November 19, 2003 7:32
Subject: [Histonet] Frozen section fixing
problems - but paraffin works!
I am working with
a new monoclonal antibody in parathyroid tissue, and I use the ABC-DAB
reageant system for immunohistochemistry.
I am new to frozen
sections immunostaining - and I am encountering problems with frozen sections
- staining is very weak or absent, in comparison to paraffin embedded tissue,
which I have no problems with.
My protocol for frozen
slides with 5 micron sections thawed at room temperature for 5
Placed in 70%
ethanol x 5 minutes.
Washed in PBS x 5
standard protocol: hydrogen peroxide 0.3% x 30 min, donkey serum 5% x 30
min, primary antibody (mouse) 4 deg overnight, secondary (goat anti-mouse),
Does anyone have
email message, including any attachments, is for the sole use of the
intended recipient(s) and may contain confidential information. Any
unauthorized review, use, disclosure or distribution is prohibited. If
you are not the intended recipient(s) please contact the sender by reply email
and destroy all copies of the original message. Thank