Hi together,
I have a question about Kim1 (kidney injury marker) and the way it appears under the microscope.
We made some test staining with this a.b.on formalin fixed/paraffinated rat kidney which worked fine in my opinion, but I was told, that the specific staining should only appear in proximal tubules(p.t) of the Cortex area(staining in Cytoplasm). Poor for me- I also have a staining in the ,,Outer stripe of the outer medulla’’(brushy boarder of p.t.), which makes the evaluation very difficult.
For the establishment of the a.b. we used kidney from 1 animal without treatment, and one treated with Gentamicine, to induce an injury of (p.t).
Poor again.... I also have a good staining in the ,,Outer stripe of the outer medulla’ even in the untreatet rat.
We use a goat anti rat a.b. by ,,R&D Systems’’ and tried several ways of blocking and pretreatment(MW, Pepsin, Trypsin, Citra,) and we even tried Alakaline Phospatase versus Peroxidase staining systems. The results looked nearly similar.
So- here are the questions:
Does someone of you know, how kim1 should look like (picture source eventually?)
Is it possible, that the brushy boarder of proximal tubules, is stained similar like spots of injured p.t.’s in the Cortex.
I’m not shure if it’s allowed to post a link, but I did not know how to upload pictures in my thread .I posted a ZIP file with 4 pictures over there
http://www.2shared.com/file/L_-QGjny/KIM1.htmlThank you very much in advance for your helpful comments and greetings from the very rainy summer in Germany.
