Antibody stuff

William M. Saidel wmsaidel at wam.umd.edu
Wed Aug 5 09:47:01 EST 1992


In article <13824 at gazette.bcm.tmc.edu> rchafetz at cephalo.neusc.bcm.tmc.edu (Ross S. Chafetz) writes:
>
>
>Question
>
>	I'm doing some antibody staining, using ABC
>(vector kit).  The sections (35um) are free floating in wells.  
? What is the fluid they are floating in? If buffer, how long?
>Lately
>my sections are sticking to themselves and to other sections (there are 7
>per well, mice brains so they are pretty small).  I'm trying to pry stuck
>sections apart with forcepts but that is also tearing the tissue.  If
>anyone else has come across this problem, what did you do?  And I do
>not want to do the staining on slides.
>
>Thanks
>
>Ross Chafetz
>
>rchafetz at cephalo.neusc.bcm.tmc.edu  
Sticking sections, in my experience, is due to i) poor fixation and/or
ii) mold in the storage medium. Tell us more about the conditions you
are working in. 1) storage medium, 2) temperature of storage, 3) nature of 
fixation.
I have stored sections in buffer after 4%paraform in 0.1M phosphate for 2
months before doing antibody staining with ABC kit with no problem (fish and	
Chicken brains).

Bill Saidel (ws2 at umail.umd.edu until 8/28!)



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