Method for detecting lymphocyte subgroup with mono-clone antibody SPA hematid rosette method
A monoclonal antibody and lymphocyte technology, applied in the field of medical immunology, can solve problems such as loss, difficulty, and inability to adapt, and achieve the effect of reducing systematic errors and accidental errors, avoiding time-consuming and complicated operations, and reducing cell damage
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Embodiment 1
[0021] Example 1 Lyophilized antibody sensitized erythrocyte rosette reagent WuT 3 、WuT 4 、WuT 8 (The freeze-dried product is a small amount of solid crystals, which dissolve after adding liquid, without affecting the volume). Add 0.5mL of calcium-magnesium-free Hank'S solution with a pH value of 7.0 to dissolve into a sensitized erythrocyte preservation solution. If the dissolved cell suspension cannot be used up, add 0.1% NaN 3 , sealed and stored at 4-6°C for 2-4 weeks.
[0022] Anticoagulate with heparin, extract 1mL of fasting venous blood from the patient, add 1mL of calcium- and magnesium-free Hank'S solution with a pH value of 7.0, mix well, and slowly add it to 2mL of lymphocyte separation medium with a dropper. Centrifuge at 800 rpm for 20 min in a 22 cm centrifuge.
[0023]Use a dropper to suck out the mononuclear cells and put them into another centrifuge tube with 4 to 6 times the amount of calcium- and magnesium-free Hank'S solution with a pH value of 7.0. As...
Embodiment 2
[0029] Example 2 Lyophilized antibody sensitized erythrocyte rosette reagent WuT 3 、WuT 4 、WuT 8 Add 0.5mL calcium-free and magnesium-free Hank'S solution with a pH value of 7.2 to dissolve into sensitized erythrocyte preservation solution. If the dissolved cell suspension cannot be used up, add 0.1% NaN 3 , sealed and stored at 4-6°C for 2-4 weeks.
[0030] Disodium salt of ethylenediaminetetraacetic acid (EDTA-Na 2 ) anticoagulant, draw 1.5mL of fasting venous blood from the patient, add 3mL of calcium- and magnesium-free Hank'S solution with a pH value of 7.2, mix well, and slowly add it to 2.5mL of lymphocyte separation solution with a dropper, and at 22°C, use A centrifuge with a radius of 22 cm was centrifuged at 1000 rpm for 18 min.
[0031] Aspirate the mononuclear cells with a dropper, put them into another centrifuge tube with 4 to 6 times the amount of calcium- and magnesium-free Hank'S solution with a pH value of 7.2, and suck as little lymphocyte separation s...
Embodiment 3
[0037] Example 3 Lyophilized antibody sensitized erythrocyte rosette reagent WuT 3 、WuT 4 、WuT 8 Add 0.5mL calcium-free and magnesium-free Hank'S solution with a pH value of 7.4 to dissolve into the sensitized red blood cell preservation solution. If the dissolved cell suspension cannot be used up, add 0.1% NaN 3 , sealed and stored at 4-6°C for 2-4 weeks.
[0038] Anticoagulate with heparin, extract 2 mL of fasting venous blood from the patient, add 4 mL of calcium- and magnesium-free Hank'S solution with a pH value of 7.4, mix well, and slowly add it to 3 mL of lymphocyte separation liquid with a dropper. Centrifuge at 1200rpm for 25min in a 22cm centrifuge.
[0039] Aspirate the mononuclear cells with a dropper, put them into another centrifuge tube with 4 to 6 times the amount of calcium- and magnesium-free Hank'S solution with a pH value of 7.4, and suck as little lymphocyte separation solution as possible. Mix well, centrifuge at 600 rpm, centrifuge for 6 minutes, d...
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