The following protocol has been developed and optimized by R&D Systems IHC/ICC laboratory for ICC experiments using cells grown on gelatin-coated glass coverslips.
This protocol provides a basic guide for the preparation, fixation, and fluorescent staining of cells on glass coverslips. 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 the staining 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 the protocol in its entirety before starting.
Many cultured cell types do not adhere well to glass coverslips. Adding a thin layer of gelatin to a coverslip enhances the adhesion of cultured cells to glass.
This protocol has been developed and optimized using R&D Systems NorthernLights fluorescent secondary antibodies but can be modified accordingly.
Note: This protocol is optimized for cells grown on coverslips in a 6- or 24-well plate but can be adapted accordingly.
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