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Hi Florian!
         I routinely spin my diluted secondary before applying to my
slides. 2 minutes in a microfuge at full speed helps to reduce the spotty
background (i.e., aggregates).
         Kathy





At 07:22 PM 3/10/04 +0100, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>--
>dear all,
>
>i am currently doing antibody stainings on paraffin sections.
>unfortunately, i am always getting very bright dots when i look at
>the staining under the confocal.when i do the control without first
>antibody, i also see these dots. therefore, they should clearly come
>from the 2nd antibody. it does not matter whether i use an alexa 488
>or alexa 546 coupled antibody, they both produce these effects. maybe
>the antibodies are bad, but they are from a commercial source and
>work fine in many applications. so, is this a probelm related to
>paraffin sectioning?
>
>i would appreciate any ideas or experience how to get around that
>problem and be forever grateful!
>
>
>best wishes,
>
>Florian
>----------------------------------------------------------------------------------------------
>Florian Ulrich
>Heisenberg Lab
>Max-Planck-Institute for Molecular Cell Biology and Genetics
>Pfotenhauerstrasse 108
>01307 Dresden
>Germany
>phone: (+49) 351 210 2689
>fax:    (+49) 351 210 1489
>email: [log in to unmask]

Kathy Spencer, Ph.D.
Scientific Associate
The Scripps Research Institute
10550 N. Torrey Pines Road
ICND 202
La Jolla, CA 92037
858-784-8437