Labeling Murine T Helper Lymphocytes using Radiolabeled Monoclonal Antibodies

For chicken ovalbumin-specific TH1 cell radiolabeling, first use a dose calibrator to draw 37 megabecquerels of the T cell-specific radioactive antibody of interest, into a syringe without dead volume.

Next, dispense the antibody into a reaction cup, and measure and subtract the remaining amount of reactivity in the syringe from the amount that was drawn. Then, add one milliliter of saline to the cup to generate a 37-megabecquerel-per-milliliter solution.

Next, add two times 10 to the sixth OVA-TH1 cells in 0.5ml of complete medium to each well of a 48-well plate, followed by 20 microliters of the radiolabeled antibody solution. After 30 minutes in a radiation-safe 37-degrees Celsius and 7.5% carbon dioxide cell culture incubator, pool the radiolabeled cells in a 50-milliliter conical tube for two washes in 10 milliliters of 37 degrees Celsius PBS.