Frozen samples were fixed in paraformaldehyde and cut into 8 μm sections; then, antigen was recovered and blocked and incubated with the primary antibodies overnight at 4°C. Afterward, we observed reaction products by secondary antibodies, and the sections were counter-stained with DAPI for 10 minutes. The following primary antibodies were used: rabbit anti-rat TH (Abcam, ab112, 1 : 100) was used to stain noradrenergic nerve fibers, and cholinergic cell bodies and nerve fibers were stained using goat anti-rat CHAT (Novus, NBP1-30052, 1 : 100). And mouse anti-rat GAP43 (Abcam, ab129990, 1 : 100) was used to localize sympathetic regeneration in the heart. The secondary antibodies used were as follows: donkey anti-rabbit IgG H&L (Abcam, ab150075, 1 : 200), donkey anti-goat IgG H&L (Abcam, ab6881, 1 : 500), and donkey anti-mouse IgG H&L (Abcam, ab150105, 1 : 600). The stained slides were observed under a Zeiss Vert A1 fluorescence microscope (Carl Zeiss Jena, German) [20 ]. Images were obtained in ten random fields per section, and analysis was performed with Image-Pro Plus analysis software (Media Cybernetics, USA).
Ab129990
Manufactured by Abcam
Ab129990 is a lab equipment product from Abcam. It is a device designed for use in scientific research applications. The core function of this product is to facilitate specific laboratory procedures, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Ab150105, by Abcam (1 mentions)
Vert a1 fluorescence microscope, by Zeiss (1 mentions)
Ab6881, by Abcam (1 mentions)
Ab150075, by Abcam (1 mentions)
Image pro plus analysis software, by Media Cybernetics (1 mentions)
Ab112, by Abcam (1 mentions)
Ab140495, by Abcam (1 mentions)
Ab205719, by Abcam (1 mentions)
Ab205718, by Abcam (1 mentions)
2 protocols using ab129990
1
Immunohistochemical Analysis of Sympathetic Nerve Fibers
2
Immunohistochemical Staining of GAP43 and 5-HT in Spinal Cord
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Immunohistochemical staining was performed according to the manufacturer’s
instructions. Briefly, paraffin-embedded spinal cord tissues were cut into 4-µm
sections and deparaffinized with xylene. Then, antigen retrieval with 3%
methanol and hydrogen peroxide was performed at room temperature for 10 minutes,
and the sections were incubated with goat serum at room temperature for 20
minutes. The sections were incubated with mouse anti-GAP43 (ab129990; 1:500;
Abcam) or rabbit anti-5-HT (ab140495; 1:1000; Abcam) at 4°C overnight and then
incubated with goat anti-mouse IgG (ab205719; 1:5000; Abcam) or goat anti-rabbit
IgG (ab205718; 1:5000; Abcam), respectively, for 30 minutes at 37°C. Then, the
sections were incubated with diaminobenzidine (Boster, Wuhan, China) for 2
minutes and counterstained with hematoxylin. Positive immunostaining was defined
as brown or yellow granules in the cytoplasm or nucleus. PBS was used instead of
a primary antibody in the negative control group. Five visual fields were
randomly selected and assessed for immunoreactive areas at 400× magnification
using the BA200 Digital image system. The images were analyzed using Image-Pro
Plus software.
instructions. Briefly, paraffin-embedded spinal cord tissues were cut into 4-µm
sections and deparaffinized with xylene. Then, antigen retrieval with 3%
methanol and hydrogen peroxide was performed at room temperature for 10 minutes,
and the sections were incubated with goat serum at room temperature for 20
minutes. The sections were incubated with mouse anti-GAP43 (ab129990; 1:500;
Abcam) or rabbit anti-5-HT (ab140495; 1:1000; Abcam) at 4°C overnight and then
incubated with goat anti-mouse IgG (ab205719; 1:5000; Abcam) or goat anti-rabbit
IgG (ab205718; 1:5000; Abcam), respectively, for 30 minutes at 37°C. Then, the
sections were incubated with diaminobenzidine (Boster, Wuhan, China) for 2
minutes and counterstained with hematoxylin. Positive immunostaining was defined
as brown or yellow granules in the cytoplasm or nucleus. PBS was used instead of
a primary antibody in the negative control group. Five visual fields were
randomly selected and assessed for immunoreactive areas at 400× magnification
using the BA200 Digital image system. The images were analyzed using Image-Pro
Plus software.
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