Isolated vagal neurons from DiI-labeled SD rats were first fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) for 10 min. After PBS washing, neurons were permeabilized in 0.1% Triton X-100 for 10 min, washed with PBS, and blocked with PBS containing 5% bovine serum albumin (BSA) for 1 h at room temperature. Next, the neurons were incubated with a mixture of rabbit anti-Dectin-1 antibody (1:50; Abcam, Cambridge, UK) and mouse anti-TRPV1 antibody (1:100; Abcam) overnight at 4 °C, which were diluted in PBS containing 5% BSA. After three washes with PBS, the neurons were incubated with a mixture of Alexa Fluor® 405-conjugated goat anti-rabbit IgG (1:200; Abcam) and Alexa Fluor® 488-conjugated goat anti-mouse IgG (1:200; Abcam) for 1 h at room temperature. The neurons were rewashed by PBS and mounted in Aqueous Mounting Medium (Abcam). Images were captured using a confocal spectral microscope (TCS SP5; Leica Microsystems, Mannheim, Germany) with a 20× objective.
Aqueous mounting medium
Manufactured by Abcam
Aqueous Mounting Medium is a laboratory product designed to mount and preserve biological specimens on microscope slides. It is an aqueous-based solution formulated to maintain the clarity and structure of samples during microscopic analysis.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 700 confocal microscope, by Zeiss (2 mentions)
Alexa fluor 488 conjugated goat anti mouse igg, by Abcam (1 mentions)
Tcs sp5, by Leica (1 mentions)
Phalloidin ifluor 488, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Rabbit anti dnase 1l1 antibody, by Atlas Antibodies (1 mentions)
2 protocols using aqueous mounting medium
1
Immunocytochemical Analysis of Vagal Neurons
2
Quantifying Macrophage DNase 1L1 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNase 1L1 staining was performed on macrophages polarized for 6 or 24 hours. Samples were blocked with 2% BSA, 0.5% fish gelatine, and 0.3% Tween20 in PBS (Morphisto) and afterwards incubated with a rabbit anti-DNase 1L1 antibody (0.2 µg/mL, Atlas Antibodies). Detection was performed using an anti-rabbit Cy3 antibody (2.5 µg/mL, Biolegend). Actin cytoskeleton as well as nuclei were stained with phalloidin-iFluor488 (1:1000, Abcam) and DAPI (1 µg/mL, Sigma-Aldrich). The slides were mounted with Aqueous Mounting Medium (Abcam). Fluorescence images were acquired on a Zeiss LSM 700 confocal microscope using a Zeiss 63x oil objective and a pinhole of 1 airy unit (≈0.8 µm slice thickness).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!