[Histonet] Dako CSAII vs Elite ABC method, which is more sensitive?

Kelly D Mcqueeney kelly.mcqueeney <@t> bms.com
Mon Mar 21 15:24:59 CST 2005


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 Envison 
+ 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 increased 
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 I 
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% Tween-20 (PBT), nickel-DAB from Vector.

Thanks,
Kelly
Bristol-Myers Squibb




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