AW: [Histonet] Antibody deposit visualization

[Gudrun Lang]

Hi,
 I think this is comparable with IF for detecting human autoimmun antibodies
in skin or kidney.

The human immunglobulins are detected with an fluorochrome conjugated
antibody mouse-anti-human IgG (for example).
In your case it may be successful to stain with a goat-anti-xxx IgG / IgM
/IgA etc.

On human FFPE kidney we performed pretreatment with pepsin (3-5 min) and
then 30 min polyclonal anti-serum 1:10 and 1:20.

To show immunglobulin-deposits in the glomeruli you can perform a
trichrome-stain like MSB or SFOG. Protein deposits will be shown bright red,
but are not distinguishable in the kind of protein.

It will depend on the amount of deposit, I assume.

Hope this helps
Gudrun


-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Rodriguez,
Kristofer
Gesendet: Mittwoch, 22. Oktober 2014 17:31
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] Antibody deposit visualization

One of my clients has been performing experiments with a mouse model of
Dengue Virus Infection. They have been treating the mice with monoclonal
antibodies (client did not specify what antibody)as a potential treatment
for dengue virus infection. They have harvested the kidneys and put them in
10% NBF. They want to know if routine H&E will show any antibody deposits.

I told him that I would not be able to directly observe the antibody deposit
itself, however I might be able to observe the reaction to the antibody
deposit in the basement membrane of the glomeruli as a membranous
glomerulopathy on an H&E.

I was wondering if I should do any special stains that might show more than
an H&E.
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