Autofluorescence

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From:"Michael J. Lyon, Ph.D." <lyonm@VAX.CS.HSCSYR.EDU> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Hello:

I have just started doing immunostaining of muscle dissections.  The tissue
has been immersion fixed in 4% paraformaldehyde 0.2% Picric acid for about
24 hrs at 4C.  My primary antibodies are all fairly dilute at 1:3000 or
1:4000, secondary is at 1:400 and I am using CY3 at 1:600.  I have tried
ammonium chloride and sodium borohydride but still have considerable
background from the muscle fibers.  I was thinking of switching to a
different fixation.  Any suggestions would be appreciated.  Hope I have
given sufficient info.

Thanks in advance

Michael




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