CART staining. 2 h food intake and stomach content were weighed upon sacrifice. NG were collected, post-fixed in 4% PFA 2 h, and cryoprotected in 0.1 M phosphate buffer containing 25% sucrose at 4°C until cryo-sectioned (10 μm). NG sections from left and right nodose ganglia of one fed and one fasted animal were placed on the same slide (1:5 series) to allow direct comparison. Slides were incubated overnight with CART antibody (1:1000; H003–62; Phoenix Pharmaceuticals) and stained 2 h with donkey anti-rabbit Ig conjugated to AlexaFluor488 (1:500; ThermoFisher Scientific). Specificity of immunostaining was determined by omitting the primary antibody or by pre-incubation of antibody with an excess of CART peptide. Stained sections were mounted on slides and coverslipped with ProLong Gold (Invitrogen) prior to imaging on Zeiss Axio Imager 2 with AxioCam503 using a 20x or 40x objective.
H003 62
Manufactured by Phoenix Pharmaceuticals
H003–62 is a laboratory equipment designed for sample preparation and analysis. It is a compact and versatile device that can be used for a variety of applications in research and testing environments.
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3 protocols using h003 62
1
Quantification of CART Expression in Nodose Ganglia
2
Retinal Cell Identification Protocol
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After recording, retinas were fixed (4% paraformaldehyde, 30 min, 20°C) and counterstained with one or more antibodies: (1) rabbit anti-cocaine and amphetamine-regulated transcript (CART; 1:1000; H00362, Phoenix Pharmaceuticals), a specific marker for most ON-OFF DS cells; Kay et al., 2011 (link)); (2) guinea pig anti-RNA-binding protein with multiple splicing (RBPMS; 1:1000; 1832-RBPMS, PhosphoSolutions), a pan-ganglion-cell marker; Rodriguez et al., 2014 (link)); (3) goat anti-ChAT (1:100, anti-ChAT; AB144P, Millipore); (4) rat anti-mCherry (1:1000, EST202, Kerafast); or (5) rabbit anti-melanopsin (1:1000, Advanced Targeting Systems).
3
Quantification of CART Expression in Nodose Ganglia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CART staining. 2 h food intake and stomach content were weighed upon sacrifice. NG were collected, post-fixed in 4% PFA 2 h, and cryoprotected in 0.1 M phosphate buffer containing 25% sucrose at 4°C until cryo-sectioned (10 μm). NG sections from left and right nodose ganglia of one fed and one fasted animal were placed on the same slide (1:5 series) to allow direct comparison. Slides were incubated overnight with CART antibody (1:1000; H003–62; Phoenix Pharmaceuticals) and stained 2 h with donkey anti-rabbit Ig conjugated to AlexaFluor488 (1:500; ThermoFisher Scientific). Specificity of immunostaining was determined by omitting the primary antibody or by pre-incubation of antibody with an excess of CART peptide. Stained sections were mounted on slides and coverslipped with ProLong Gold (Invitrogen) prior to imaging on Zeiss Axio Imager 2 with AxioCam503 using a 20x or 40x objective.
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