Treating TEM-cuts with whole-mount gold AB

Jens Krieger krieger at mibm3.ruf.uni-freiburg.de
Mon Oct 7 09:35:09 EST 1996


Hello!
I've a problem with my ultrathin cuts for AB-staining treating them with
IgG-whole gold mount AB from Sigma. Before I use the AB I treated them
with 1% BSA in PBS+TWEEN 20 for about half an hour. Then I use my first AB
in PBS+TWEEN for three hours at roomtemperatur. Washing three times in
PBS+TWEEN, incubating with the gold in PBS+TWEEN for about two hours at
RT, drying and.....sh....There is a very high background on my grids. Like
the BSA falls out or anything else. I can't explain...
Does anyone know why there is such a high background? Or does anyone has a
hint how a can improvet the immunolabeling of my cuts?
Thanks a lot in advance,
			Jens.

/~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\
|  Jens Krieger		"Solange die Alpen noch stehen, |
|  				wird es keine		|
|  Tel.:+49.351.463.5465  	   Gerechtigkeit geben."|
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|  krieger at rcs.urz.tu-dresden.de	H.Achternbusch	|
\  krieger at mibm.ruf.uni-freiburg.de			/
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