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Encyclopedia of Experiments

A Fluorescence-Based Assay Using Engineered Lymphocytes for Screening Immunomodulatory Compounds

This video demonstrates the screening of immunomodulatory compounds via fluorescent-based assay. Transgenic mouse-derived T lymphocytes with green fluorescent protein expression cassettes are treated with these compounds. The enhanced and reduced GFP signals enable the identification of the compounds' stimulatory and inhibitory effects, respectively.

1. Preparation of Splenocyte Medium and Flow-cytometry Buffer

  1. Perform all steps under a 70% ethanol-cleaned biological hood.
  2. Remove 70 mL of pre-warmed Roswell Park Memorial Institute (RPMI) 1640 1x medium (commercially available and supplied in filtered 500 ml volume sterile bottles) and place in a sterile tube. The removed medium will be used later in the protocol.
  3. Supplement the remaining 430 mL of RPMI 1640 1x with 50 ml inactivated fetal bovine serum (FBS), 5 mL penic.......

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