After 14 and 28 d, samples were fixed with 4% paraformaldehyde, embedded in paraffin, and cut into 5-μm sections. Sections were stained with hematoxylin and eosin (H&E) for general histology, and Alcian blue and Safranin O staining were performed to evaluate chondrogenesis. Expressions of type II collagen (COL II), type I collagen (COL I), and aggrecan were examined by immunohistochemical staining. Paraffin‐embedded sections were deparaffinized and dehydrated. Primary antibodies were incubated overnight in 4 °C. The SABC immunohistochemical kit (SA1020, Boster, Wuhan, China) was used and DAB (AR1022, Boster, Wuhan, China) was used as the chromonogen. Stained sections were scanned using a digital pathological scanning system (Aperio CS2, Leica, Germany) at 200 magnification.
Sabc immunohistochemical kit
Manufactured by Boster Bio
The SABC immunohistochemical kit is a laboratory equipment used for the detection and visualization of target proteins or antigens in tissue samples. It utilizes the streptavidin-biotin complex (SABC) method to amplify the signal and enhance the sensitivity of the immunohistochemical staining process.
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2 protocols using sabc immunohistochemical kit
1
Chondrogenesis Evaluation in Tissue Sections
2
Optimized Immunohistochemistry Protocol
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For immunohistochemistry, 5 μm cryosections were collected on adhesive slides and fixed in 4% PFA for 10 min, then the slides were washed with PBS 3 times and then antigen-retrieved with sodium citrate using heating method with microwave. Slides were washed in PBS for 3 times and blocked in 5% BSA for 1 h at 37°C, then incubated at 4°C overnight with primary antibodies. The slides were stained following the SABC immunohistochemical kit (BOSTER) protocol, and nuclei were visualized by hematoxylin stain and observed using Nikon NID-elements software.
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