Immunohistochemistry and Immunofluorescence Assays
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Corresponding Organization : Medicon (United States)
Variable analysis
- Cell permeabilization with 0.1% Triton X-100 in PBS for 15 min
- Incubation with primary antibodies at 4 °C overnight
- Incubation with horseradish peroxidase-conjugated secondary antibody at room temperature for 1 h
- Development with chromogen (DAB) for 5–10 min at room temperature
- Immunohistochemical staining of cells
- Immunofluorescence detection of cells
- Cells transferred onto glass slides
- Washing cells with PBST (PBS + 0.05% Tween 20) 3 times
- Blocking cells with normal serum or 5% bovine serum albumin for 1 h
- Washing cells with PBST for 15 min after incubation with primary and secondary antibodies
- Counterstaining cells with hematoxylin
- Dehydrating and mounting slides with liquid mounting media
- Covering fluorescence-stained slides with mounting medium containing DAPI
- Examining and photographing stained slides using a Nikon fluorescence microscope
- Not explicitly mentioned
- Not explicitly mentioned
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