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Carbonyl staining question


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#1 Triwinters

Triwinters

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Posted 21 July 2010 - 01:20 AM

I am trying to label carbonyls to measure oxidative stress for immunofluorescence on frozen sections

I had a few questions regarding this.

for a positive control would H202 work before fixing, or should I use H202 + Fe (2+)?
Also for anyone that might have experience with this, were the fluorescence increases easily distinguishable?

I have been having a problem with this and can't tell if the fluorescence is just low or if I need to tweak my protocol

General protocol
fix - methanol 50% or methanol/acetone
DNPH in HCl
block, primary, secondary.


Thanks in advance

Edited by Triwinters, 21 July 2010 - 08:23 AM.





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