After the blocking step, the cells were incubated with primary antibody-anti-vimentin (1:100, Boster, Wuhan, China), anti-keratin (1:100, Boster) at 4 °C overnight; PBS was used as the negative control. The cells were then washed with PBS and incubated for 1 h with the secondary antibodies, namely anti-mouse IgG Alexa Fluor-488 or at antirabbit IgG Alexa Fluor-594 1:1000 at room temperature (Life Technologies, Paisley, UK). Glass cover slips were mounted using mounting media supplemented with DAPI stain (VectorLabs, Peterborough, UK) and preparations imaged under a fluorescent microscope (Olympus, Tokyo, Japan).
Anti keratin
Anti-keratin is a laboratory reagent used for the detection and quantification of keratin, a structural protein found in various tissues, including skin, hair, and nails. It is commonly used in immunohistochemistry, Western blotting, and other analytical techniques to study the expression and distribution of keratin in biological samples.
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2 protocols using anti keratin
Immunofluorescence Staining of HDPFs
After the blocking step, the cells were incubated with primary antibody-anti-vimentin (1:100, Boster, Wuhan, China), anti-keratin (1:100, Boster) at 4 °C overnight; PBS was used as the negative control. The cells were then washed with PBS and incubated for 1 h with the secondary antibodies, namely anti-mouse IgG Alexa Fluor-488 or at antirabbit IgG Alexa Fluor-594 1:1000 at room temperature (Life Technologies, Paisley, UK). Glass cover slips were mounted using mounting media supplemented with DAPI stain (VectorLabs, Peterborough, UK) and preparations imaged under a fluorescent microscope (Olympus, Tokyo, Japan).
Immunofluorescence Assay for Podocyte Characterization
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