From owner-cellbiol@net.bio.net Tue Jan 14 22:00:00 1997
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From: gilbride@bu.edu (Kevin Gilbride)
Newsgroups: bionet.cellbiol
Subject: Re: lysis buffer for monolayer culture + ELISA
Date: Wed, 15 Jan 1997 12:35:58 -0500
Organization: Mallory Institute of Pathology
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Message-ID: <gilbride-1501971235580001@ppp-83-24.bu.edu>
References: <32B7FE9A.351A@rzmail.uni-erlangen.de>
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In article <32B7FE9A.351A@rzmail.uni-erlangen.de>, "Dr. O. Pullig"
<oliver.pullig@rzmail.uni-erlangen.de> wrote:

> HI,
> I´m doing monolayer cultures of chondrocytes and want to lyse the cells
> and also get the extracellular matrix in solution for quantitation of
> collagens with inhibition ELISA. Therefore the buffer should be
> - able to lyse the cells and bring also the matrix in solution
> - work with ELISA 
> - should be able get concentrated in centricons
> 
> any suggestions
> 
> Oliver


Hello Oliver,

      Try tris, deoxycholic acid, SDS with a PMSf protease inhibitor. We
used this with our Laminin ELISAs. You could also try a protease with SDS
heated to about 60 deg C for ~ 30 min. ultracentrifuge 100,000 X g and you
should be all set. 



Good Luck,

KJ Gilbride
Mallory Inst. of Path.
Boston Univ. Sch. of Med.

.