Kidney tissue was immersed in fixative containing 3.7% formaldehyde, 10 mM sodium metaperiodate, 40 mM phosphate buffer and 1% acetic acid. The tissue was dehydrated through a graded ethanol series, embedded in paraffin, sectioned (4 μm) and mounted on glass slides. For immunofluorescence (IF) staining, sections were incubated for two rounds of staining overnight at 4 °C. Anti-rabbit or anti-mouse IgG-horseradish peroxidase was used as a secondary antibody. Each round was followed by tyramide signal amplification with the appropriate fluorophore (Alexa Fluor 488 tyramide, Alexa Flour 647 tyramide or Alexa Fluor 555 tyramide, Tyramide SuperBoost Kit with Alexa Fluor Tyramides, Invitrogen) according to the manufacturer’s protocols. 4′,6-Diamidino-2-phenylindole was used as a nuclear stain. Sections were viewed and imaged with a Nikon TE300 fluorescence microscope and spot-cam digital camera (Diagnostic Instruments), followed by quantification using Image J (NIH).
Tyramide superboost kit with alexa fluor tyramides
Manufactured by Thermo Fisher Scientific
The Tyramide SuperBoost Kit with Alexa Fluor Tyramides is a laboratory equipment product designed to enhance fluorescent signal detection in various biological applications. It utilizes a tyramide-based amplification system to increase the intensity of fluorescent signals, enabling improved visualization and analysis of target analytes.
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2 protocols using tyramide superboost kit with alexa fluor tyramides
1
Immunofluorescence Staining of Kidney Tissue
2
Immunofluorescence Staining of Kidney Tissue
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Kidney tissue was immersed in fixative containing 3.7% formaldehyde, 10 mM sodium m-periodate, 40 mM phosphate buffer, and 1% acetic acid. The tissue was dehydrated through a graded series of ethanols, embedded in paraffin, sectioned (4 μm), and mounted on glass slides. For immunofluorescence staining, the sections were incubated for two rounds of staining overnight at 4°C. Anti-rabbit or mouse IgG-HRP was used as a secondary antibody. Each round was followed by tyramide signal amplification with the appropriate fluorophore (Alexa Fluor 488 tyramide, Alexa Flour 647 tyramide or Alexa Fluor 555 tyramide, Tyramide SuperBoost Kit with Alexa Fluor Tyramides, Invitrogen) according to the manufacturer’s protocols. DAPI was used as a nuclear stain. Sections were viewed and imaged with a Nikon TE300 fluorescence microscope and spot-cam digital camera (Diagnostic Instruments), followed by quantification using Image J software (NIH, Bethesda, MD).
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