AW: [Histonet] Antibody deposit visualization
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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Rodriguez, Kristofer Gesendet: Mittwoch, 22. Oktober 2014 17:31 An: histonet@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 HE 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 HE. I was wondering if I should do any special stains that might show more than an HE. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[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 HE 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 HE. I was wondering if I should do any special stains that might show more than an HE. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Antibody deposit visualization
Depending on the reaction of the glomerulii to the Dengue virus and the stage of such reaction perhaps a PAM (periodic acid methenamine silver) stain could show something.René J. On Wednesday, October 22, 2014 11:31 AM, Rodriguez, Kristofer kristofer.rodrig...@bcm.edu wrote: 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 HE 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 HE. I was wondering if I should do any special stains that might show more than an HE. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Antibody deposit visualization
Hi, may be you can try to label murine immunoglobulins. A mere secondary against murine IgGs could do the trick, depending on which isotype your client is suspecting to form the precipitates. Just an idea, never have tried before. Cheers Julio Rodriguez, Kristofer kristofer.rodrig...@bcm.edu escribió: 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 HE 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 HE. I was wondering if I should do any special stains that might show more than an HE. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet