[Histonet] Immuno Bed Plastic Question

[Allyse Mazzarelli]

Hi histonetters!

I have posted on this forum a while ago asking for opinions regardnig new
types of plastics that work well with IHC. It was brought to my attention
that "Immuno-Bed" (Polysciences) works rather well. I recently purchased
the plastic, infiltrated/embedded well, and sectioned great, BUT, when I
removed the excess GMA/rehydrated and labeled with my antibodies, NOTHING
WORKED! Nothing showed up! Only DAPI!! The mechanism I used for removing
GMA and rehydrating were as follows:

1.) 3 changes of 30 minutes each in 60 Celsius xylene. (Note: I recently
let the slides sit in 60 Celsius xylene overnight too... but it did not
work!!)
2.) 3 changes of 10 minutes each in 100% EtOH.
3.) 15 minutes in 95%, 75% & 50% EtOH
4.) 5 minutes in water
5.) 3 changes of 5 minutes each, PBS.

I continued to block and apply my antibodies as normal. (Antibodies are all
brand new, not duds!)

Since this plastic was formulated specifically for IHC, I am at a loss. If
anyone has used this plastic before, any insight directed my way would be
extremely helpful!!

Thanks,

Allyse Mazzarelli
Histologist
NEUROTECH USA INC.