The collected kidneys were fixed in 10% formalin, dehydrated in a graded series of alcohol, cleaned with xylene, embedded in paraffin, and sectioned at a thickness of 5 μm. Periodic acid-Schiff and Masson staining was performed to facilitate the examination of the glomerular basement membrane and mesangial matrix of the kidneys. Immunohistochemistry was used to assess the expression of TNF-α (Abcam, ab220210), IL-1β (Cell Signaling Technologies, #12242), F4/80 (Abcam, ab100790), FN (Santa Cruz Biotechnology, SC-8422), and collagen I (Abcam, ab253113) in the kidney sections. After 3,3’-diaminobenzidine staining, cells that were positive for target protein expression stained brown-yellow, whereas the cells without the said expression were unstained. The percentage of each area that was stained was quantified using the Image-Pro Plus software.
Sc 8422
Manufactured by Abcam
SC-8422 is a lab equipment product. It serves a core function, but a detailed and unbiased description cannot be provided without extrapolation or interpretation.
Automatically generated - may contain errors
2 protocols using sc 8422
1
Histopathological Analysis of Kidney Tissue
2
Multimodal Visualization of Extracellular Matrix
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ExAdEx samples were fixed with 4% PAF and then incubated with primary anti-collagen 1 (Abcam ab260043), anti–elastin (Abcam ab21610), anti-laminin (ab11575), anti-fibronectin Santa Cruz sc8422) and anti-CD31 (Abcam ab28364) antibodies overnight at 4 °C and then with the corresponding secondary antibody for 45 min at room temperature. Lipid droplets were stained with Oil Red O (ORO), and nuclei were stained with DAPI. Samples were visualized on an LSM 780 NLO inverted Axio Observer Z1 confocal microscope (Carl Zeiss Microscopy GmbH, Jena, Germany) using a Plan Apo 25× multi immersion (oil, glycerol, water) NA 0.8 objective. The SHG light source was a Mai Tai DeepSee (Newport Corp., Irvine, CA, USA) tuned to 880 nm. A forward SHG signal was detected with an oil condenser (1.4 NA), bandpass filter 440/40 nm and transmission PMT. Backward SHG was collected with a GaAsP (BIG) nondescanned module of 440/10 nm.
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