The femoral tissues were routinely sectioned, and the baked flakes were dewaxed with xylene and hydrated sequentially with a gradient ethanol solution. The antigen was repaired in citrate buffer for 20 min, inactivated in 3% hydrogen peroxide solution for 30 min, and blocked with 5% bovine serum albumin (BSA) for 20 min. The primary antibodies rabbit anti-OPG-antibody (1:50, ab73400, Abcam) and anti-RANKL antibody (1:100, sc52950, Santa Cruz Biotechnology, USA) were added dropwise and incubated at 4 °C overnight. After rewarming, the sections were incubated with horseradish peroxidase-labeled goat anti-rabbit IgG and anti-mouse IgG (Abcam, UK) at 37 °C for 1 h. DAB (Solarbio, Beijing, China) was used for colour development, and the sections were lightly counterstained, dehydrated, made transparent, and sealed. Three sections of each tissue were observed under a 100× optical microscope (Olympus, Japan) to analyse the positive cells.
Horseradish peroxidase labeled goat anti rabbit igg and anti mouse igg
Manufactured by Abcam
Sourced in United Kingdom
Horseradish peroxidase-labeled goat anti-rabbit IgG and anti-mouse IgG are secondary antibodies used in various immunoassays and detection techniques. They are conjugated with the enzyme horseradish peroxidase, which can catalyze a colorimetric or chemiluminescent reaction for visualization and quantification purposes.
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2 protocols using horseradish peroxidase labeled goat anti rabbit igg and anti mouse igg
1
Immunohistochemical analysis of OPG and RANKL
2
Femoral Tissue Immunohistochemistry
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The femoral tissue were routine sectioned, the baked akes were dewaxed with xylene and hydrated sequentially with a gradient ethanol solution. The antigen was repaired in citrate buffer for 20 min, 3% hydrogen peroxide solution was inactivated for 30 min, and blocked with 5% BSA for 20 min. The anticalcitonin receptor (CALCR) polyclonal antibody (1:20, ab11042, Abcam, UK) and anti-cathepsin K (CTSK) polyclonal antibody (1:1000, ab19027, Abcam), rabbit anti-OPG-antibody (1:50, ab73400, Abcam) and anti-RANKL antibody (1:100, sc52950, Santa Cruz Biotechnology, USA)were added dropwise and reacted at 4 °C overnight. After rewarming, the sections were incubated with the horseradish peroxidase-labeled goat anti-rabbit IgG and anti-mouse IgG (Abcam, UK) at 37 °C for 1 h. DAB (Solarbio, Beijing, China) was used for color development, hematoxylin was lightly counterstained, dehydrated, transparent, and sealed. Three sections of each tissue were observed under a 100× optical microscope (Olympus, Japan) to analyze positive cell.
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