Canine Retinal CNGA3 Immunohistochemistry

For the ex vivo data, IHC staining was performed on 10 μm thick canine retinal cryosections that were incubated overnight at +4°C in the presence of rabbit polyclonal anti-canine CNGA3 antibody (courtesy of A. Komáromy, Michigan State University) at a concentration of 1:750. Primary antibody was visualized with Alexa Fluor 568nm goat anti-rabbit secondary antibody and cell nuclei were stained with DAPI. Slides were mounted (Gelvatol, Sigma-Aldrich) and examined by epifluorescence (Axioplan; Carl Zeiss Meditec) or confocal (Leica SP5-II, Leica) microscopy using previously published methods [31 (link)].

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Tanaka N., Dutrow E.V., Miyadera K., Delemotte L., MacDermaid C.M., Reinstein S.L., Crumley W.R., Dixon C.J., Casal M.L., Klein M.L., Aguirre G.D., Tanaka J.C, & Guziewicz K.E. (2015). Canine CNGA3 Gene Mutations Provide Novel Insights into Human Achromatopsia-Associated Channelopathies and Treatment. PLoS ONE, 10(9), e0138943.

Publication 2015

Gelvatol Axioplan Leica SP5-II

Anti antibody Antibody Canine Cell nuclei Cryosections Dapi Goat Microscopy Rabbit Retinal

Corresponding Organization : North Cumbria Integrated Care NHS Foundation Trust

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Variable analysis

independent variables

Rabbit polyclonal anti-canine CNGA3 antibody concentration (1:750)

dependent variables

CNGA3 protein expression (visualized by Alexa Fluor 568nm goat anti-rabbit secondary antibody)

control variables

Canine retinal cryosections (10 μm thick)
Incubation duration (overnight at +4°C)
Cell nuclei staining (DAPI)

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