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The following protocol has been developed and optimized by R&D Systems IHC/ICC laboratory for fluorescent IHC experiments using frozen tissue samples.
This protocol provides a basic guide for the fixation, cryostat sectioning, and staining of frozen tissue samples. Each investigator must determine the precise experimental conditions required to generate a strong and specific signal for each antigen of interest. If R&D Systems primary antibodies are employed, please refer to the product data sheets to obtain the recommended working dilutions. In this protocol, signal visualization is achieved using R&D Systems NorthernLights™ range of fluorescent secondary antibodies and reagents. For all other reagents, please follow the manufacturer’s instructions.
Please read protocol in its entirety before beginning.
This technique utilizes formaldehyde-based fixation before the tissue is frozen and sectioned using a cryostat.
This method utilizes frozen tissues that are fixed after snap-freezing and sectioning with a cryostat.
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