[Histonet] RE: Dako CSAII vs Elite ABC method, which is more sensitive?
When it comes the question which of the two techniques is more
sensitive I can predict that very certainly the CSA II kit will beat
the Elite ABC method for. It's just a matter of choices what do you
prefer and what is technically possible. Personally, I use the CSA II
kit as my last resort in case the signal with other
detection techniques is too weak:
Start using the Elite ABC kit (or EnVision, or other "simple"
detection system) and titrate your primary with appropriate
pretreatment over a wide range at a proven positive control. If it
works fine, you're done. If the signal is too weak, testing with the
CSA II kit will be step 2. Again, perform a titration experiment over
a wide range at the same positive control tissue.
You should also consider that CSA II kit signal is less crisp than
with other IHC detection systems. Usually this isn't too disturbing.
The dotty appearance of the reaction product is due to the tyramide
reaction and can therefore not be improved.
Hope this helps.
Chris van der Loos, PhD
Dept. of Pathology M2-230
Academical Medical Center
NL-1105 AZ Amsterdam, The Netherlands
----- Original Message -----
Kelly D Mcqueeney
Mon, 21 Mar 2005 16:24:59 -0500
[Histonet] Dako CSAII vs Elite ABC method, which is more sensitive?
I have been using Elite ABC kit for c-fos IHC in brain. We are getting
pretty good staining and want to improve sensitivity. I compared the
Elite ABC kit, Dako's CSA tyramide amplification kit, and Dako's
+ kit. The sections are slide-mounted, 20 micron (I increased Dako's
incubation times to 45 minutes). Dako claims that the tyramide and
Envision kit are much more sensitive than Elite ABC. I have used the
tyramide system in the past and was really impressed with the
signal. Unfortuantely, Dako's CSA and Envision kit exhibit very weak
staining and the Elite ABC kit exhibits dark and specific staining in
the brain. Has anyone else noticed a change in either product? Should
order HRP-conjugated c-fos and amplify the signal? Any suggestions
(besides floating sections)?
Method: Quench w/Dako's H2O2, block 1 hour 3% BSA in PBT, c-fos O/N,
wash buffer is PBS +! 0.01% Tw
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