RE: DAB haze

From:"Cook, Angela"

Thanks for the tips, but most of those suggestions we already use.  We have
4 rinses between steps  with a buffer containing tween.  We don't use
adhesive in the bath and we do used charged slides (Fisher Brand SuperFrost

-----Original Message-----
From: Dawson, Glen []
Sent: Thursday, August 22, 2002 4:32 PM
To: Cook, Angela
Subject: RE: DAB haze


Watch your rinses.  If you have a weak rinse between steps, this haze will
occur.  On your next run, watch the machine and see if there is an adequate
rinse between each step.  Also, you may try adding a bit of tween (soap) to
your wash buffer to get a better rinse.

Another tip would be to check the amount of adhesive there is in your
floatation bath.  If the brown haze comes up to the exact point you dip your
slides into your waterbath to pick up sections, your adhesive may be causing
the staining.  Try using a charged slide with a few antibodies with no
adhesive at all in the water bath to test this theory.

Good Luck,

Glen Dawson
Milwaukee, WI

-----Original Message-----
From: Cook, Angela []
Sent: Thursday, August 22, 2002 3:04 PM
To: HistoNet (E-mail)
Subject: DAB haze

We use a DAKO Autostainer and DAB as a chromagen.  We have been seeing a
brown haze all over the slides all the way up to the slide labels.  We have
been seeing a lot of DAB collecting in the sink of the machine as well DAKO
tells us that they don't know what is happening.  It doesn't seem to be a
messed up lot of DAB and we assayed our buffer to make sure that it had zero
protein content.  Anyone have any ideas as to what thi

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