Human DRGs were fixed in 4% paraformaldehyde overnight and then sections (12 μm) were cut on a cryostat. The sections were first blocked with 2% BSA for 1 h at room temperature, then incubated with peripherin (1:500, rabbit, Sigma) or a mixture of primary polyclonal TRPV1 antibody (1:400, rabbit, Neuromics, Edina, MN), and monoclonal NF200H antibody (1:1000, mouse, Sigma) overnight at 4°C. The sections were then incubated for 2 h at room temperature with cyanine 3 (Cy3)- and FITC-conjugated secondary antibodies (1:400; Jackson ImmunoResearch, West Grove, PA). The DRG sections were then stained with DAPI (1:1000, Sigma, for 5 min) and examined under a Nikon fluorescence microscope. Images were captured with a CCD Spot camera and analyzed with NIH Image J software or Adobe PhotoShop.
Cyanine 3 cy3 and fitc conjugated secondary antibodies
Manufactured by Jackson ImmunoResearch
Cyanine 3 (Cy3)- and FITC-conjugated secondary antibodies are fluorescent-labeled reagents used in immunodetection techniques. Cy3 is a red-orange fluorescent dye, while FITC is a green fluorescent dye. These conjugated secondary antibodies bind to primary antibodies, allowing for the visualization and detection of target proteins or molecules in various applications, such as immunohistochemistry, Western blotting, and flow cytometry.
Automatically generated - may contain errors
2 protocols using cyanine 3 cy3 and fitc conjugated secondary antibodies
1
Immunohistochemical Characterization of DRG Neurons
2
Immunohistochemical Visualization of Neuronal Markers
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Human DRGs were fixed in 4% paraformaldehyde overnight and then sections (12 μm) were cut on a cryostat. The sections were first blocked with 2% BSA for 1 h at room temperature, then incubated with peripherin (1:500, rabbit, Sigma) or a mixture of primary polyclonal TRPV1 antibody (1:400, rabbit, Neuromics, Edina, MN) and monoclonal NF200H antibody (1:1000, mouse, Sigma) overnight at 4 °C. The sections were then incubated for 2 h at room temperature with cyanine 3 (Cy3)- and FITC-conjugated secondary antibodies (1:400; Jackson ImmunoResearch, West Grove, PA). The DRG sections were then stained with DAPI (1:1000, Sigma, for 5 min) and examined under a Nikon fluorescence microscope. Images were captured with a CCD Spot camera and analyzed with NIH Image J software or Adobe PhotoShop.
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