[Histonet] Wanted dead or alive - tissue negative for connexin 43

From:"Louise Carrington"

hello ,
I am trying to find a tissue negative for connexin 43 (Cx43) to preadsorb an
antibody with and/or use as a negative staining tissue - i have been using it to
immuno-stain fresh-frozen and frozen/fixed (ethanol, acetone, 1%, 4% pfa) and
have ended up with nuclear staining in rat, human and bovine tissue. it looks
'real' ie heterogenous cell-cell staining, its not my secondary - as ommission
of primary yeilds no background (i'm using alexafluor fluorescent secondaries)
or my fixation as all fixation prtocols including no-fix show the same and i've
found it in several differnet organs (whilst searching for a positive control)
in several different species...... but the antibody is to a phosphorylated
isoform of cx43 and my my pan connexin 43 antibody shows no nuclear staining at
all. The phospho-cx43 has not been published yet and the company  have been
ignoring my emails so if anyone can suggest a tissue (or even cell culture as i
get staining here as well) that has nucleate Connexin43 cells within it that are

a) abundant and easy to isolate so that i can preadsorb the antibody OR 

b) easy to recognise so that i can do immunostaining 

(or god help me ... both??)
i would be really grateful,

i have been searching but cx 43 is a tad ubiquitous :)

thankyou in advance


Louise Carrington PhD.
Cell & Molecular Unit,
Department of Optometry & Vision Sciences,
Cardiff University,
Redwood Building,
King Edward VII Ave.,
Cathays Park,

Tel 02920 875665
email: CarringtonL@cardiff.ac.uk

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