Single-Molecule Immunofluorescence Imaging
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Corresponding Organization : Howard Hughes Medical Institute
Protocol cited in 46 other protocols
Variable analysis
- Concentration of biotinylated antibody (~10 nM)
- Incubation time of biotinylated antibody (10 min)
- Single molecule data acquired using prism type total internal reflection fluorescence (TIRF) microscope
- MPEG passivated quartz slides doped with biotin PEG
- NeutrAvidin (Thermo) coated flow chambers
- Dilution buffer (10 mM Tris-HCl pH 8.0, 50 mM NaCl, 0.1 mg/ml bovine serum albumin)
- Incubation time of fluorescent protein tag sample (20 min)
- Incubation time of prey protein antibody (~10 nM) (20 min)
- Incubation time of fluorescent-dye-labeled secondary antibody (2–5 nM) (5 min)
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