Introduction
Prepare for cell suspension
1. Isolate spleen (or lymph nodes) from 5- to 10-week-old mice, and place in 5 mL of RPMI+ in 6 cm dish.
2. Cover the tissue with a 2×2 square of sterile nylon mesh, gently smash the tissue against the mesh using the thumb side of a 5-mL syringe plunger.
3. Filtrate cell suspension with 40 μm cell strainer, wash the dish and cell strainer with 5 mL PRMI+, and collect cell suspension in a 15 mL tube.
4. Invert on ice with a few times, centrifuge the cells at 800 rpm for 5 min at 4°C, remove supernatant.
5. Add 2 mL ACK buffer per spleen, gently suspend for 1 min to lyse erythrocytes, and add 10 mL of RPMI+.
1. Isolate spleen (or lymph nodes) from 5- to 10-week-old mice, and place in 5 mL of RPMI+ in 6 cm dish.
2. Cover the tissue with a 2×2 square of sterile nylon mesh, gently smash the tissue against the mesh using the thumb side of a 5-mL syringe plunger.
3. Filtrate cell suspension with 40 μm cell strainer, wash the dish and cell strainer with 5 mL PRMI+, and collect cell suspension in a 15 mL tube.
4. Invert on ice with a few times, centrifuge the cells at 800 rpm for 5 min at 4°C, remove supernatant.
5. Add 2 mL ACK buffer per spleen, gently suspend for 1 min to lyse erythrocytes, and add 10 mL of RPMI+.
Contact Info
Address:
45-1 Ramsey Road, Shirley, NY 11967, USA
Shirley
New York 11967
United States
Shirley
New York 11967
United States
Tel:
Website:
https://www.creative-diagnostics.com/mouse-naive-cd4-t-cells-isolation.htm
Hours of operation
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