A tumor immune T cell detection kit and method
A detection kit and cell technology, applied in the field of precise biomedical detection, can solve the problems of imperfect detection targets, adverse cytokines, T cell inactivation, etc., and achieve the effects of low cost, high sensitivity, and accurate quantification.
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[0035] 1. Purification and culture of PBMC cells
[0036] Reagent:
[0037]
[0038] operate:
[0039] 1. Take 15ml of human peripheral blood, anticoagulate (EATA or heparin) treatment, room temperature for 15 minutes.
[0040] 2. Centrifuge at 350g for 5 minutes at room temperature, and draw out the plasma for later use. Add 1 volume of PBS buffer to the blood cells to dilute and mix. (If using blood components, add 1 volume of PBS buffer, dilute and mix.)
[0041] 3. Take a 15ml centrifuge tube and add 5ml of lymphocyte separation medium. Slowly add 5ml of diluted blood cells to the upper layer.
[0042] 4. Centrifuge at 450g for 20 minutes at room temperature. From top to bottom, blood cells are divided into platelet layer, white blood cell layer (buffy coat) and red blood cell layer.
[0043] 5. Aspirate the platelet layer (approximately 2ml) with a pipette. Transfer the buffy coat (buffy coat, about 3ml) to a new 15ml centrifuge tube.
[0044] 6. Add 10ml PBS ...
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