[Histonet] re: Biocare Decloaker HIER

[Linda M Watson]

Hi Jan,
We routinely use the Decloaker from Biocare. Normally, we use citrate 
buffer, pH=6.0 but that changes depending on tissue/antibody 
combination. I usually set the time around 1 minute, although I have 
gone as low at 30 seconds and as high as 3 minutes. The "set" 
temperature also varies depending on the number of containers I have. If 
I only have 1 or 2 plastic coplin jars then I set the temp at around 121 
degrees C. If I have a large Tissue Tek container with approx 15-20 each 
then the temp needs to be at around 124.5 to 126 degrees C. The trick is 
to maintain the psi at around 18 _+_ 3 or so. Basically, there are no 
rules and you have to play with the parameters to obtain the optimal 
conditions for your assay. There are other folks in my lab area that set 
it for 15 minutes at around 95 degrees C, but the psi never goes above 5 
or so. Sorry that I could not be more specific. For me, it really helps 
to understand the specificity of my antibody and the cellular expression 
/ location.

Good Luck,
Linda

Jan Shivers wrote:

>For those of you who use Biocare's Decloaker for HIER on regular cases (not prion protein retrieval), could you tell me how long you do HIER with these machines?  I normally do my prion cases in a Decloaker for 20', with a 25' cooldown, but I suspect that regular tissues don't need that length of time at high pressure/temp.  I have an extra Decloaker and am considering switching to this device for my normal case work HIER, if it's time efficient.
>
>Please reply privately.   Thanks.
>
>Jan Shivers
>Senior Scientist
>Histology/IHC/EM Section Head
>Pathology Teaching Program
>University of Minnesota
>Veterinary Diagnostic Laboratory
>1333 Gortner Ave.
>St. Paul, MN  55108
>612-624-7297
>shive003 <@t> umn.edu
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