Journal: American Journal of Physiology - Renal Physiology
Article Title: The rat kidney contains high levels of prouroguanylin (the uroguanylin precursor) but does not express GC-C (the enteric uroguanylin receptor)
doi: 10.1152/ajprenal.00282.2010
Figure Lengend Snippet: Additional features of proUgn immunostaining in kidney. A: immunoperoxidase-stained tubule labeled with antibody 6910, viewed with differential interference contrast optics. The brown reaction product is restricted to the cytoplasm of the epithelial cells that form the tubule. The nuclei of the labeled cells are in close proximity to the apical membrane. B: immunoperoxidase-stained section labeled with antibody 6910 and lightly counterstained with hematoxylin to reveal cellular features. Immunoreactive tubules are light brown, while immunonegative tubules are gray-blue. As in B, the reaction product is intracellular, and the immunopositive cells have apically oriented nuclei. C: superimposed double-label fluorescence images of kidney cortex stained with anti-proUgn antibody 6910 (blue) and anti-CD 26 antibody (red). D and E: superimposed fluorescence images of kidney cortex showing autofluorescence (green) and proUgn-like immunoreactivity (blue) as detected by anti-proUgn antibody 6910 (D) or anti-proUgn antibody 6911 (E). Peanut agglutinin (PNA) binding (F), autofluorescence (G), and proUgn-like immunoreactivity (H) are shown in a section of kidney cortex. I: superimposed images F–H. Aquaporin-2 (AQP2)-like immunoreactivity (J), autofluorescence (K), and proUgn-like immunoreactivity (L) are shown in a section of kidney cortex. M: superimposed images J–L. Scale bars = 10 μm (A), 30 μm (B–E), and 50 μm (F–M). Asterisks mark unstained glomeruli.
Article Snippet: Sections were incubated overnight at 4°C with pairs of the following reagents: a 1:50 dilution of mouse anti-CD-26 (dipeptidyl peptidase-4; DPP-IV, Thermo Scientific), a 1:1,000 dilution of rabbit anti-proUgn antibody 6910 or 6911 (raised in-house), a 1:100 dilution of goat anti-aquaporin-2 (Santa Cruz Biotechnology, Santa Cruz, CA), or a 1.66 μg/ml solution of Alexa 658-conjugated peanut agglutinin (PNA; Invitrogen).
Techniques: Immunostaining, Staining, Labeling, Fluorescence, Binding Assay