作业帮您还在做BrdU实验方法吗
来源：学生作业帮助网 编辑：作业帮 时间：2024/05/03 19:55:05
您还在做BrdU实验方法吗
您还在做BrdU实验方法吗
您还在做BrdU实验方法吗
你说的是细胞周期的流式细胞伩检测实验方法（Brdu）?
操作步骤：
1. Cells (1x106/mL) are incubated with BrdU 10 mM at final concentration, for 30 min at 37 °C in controlled atmosphere.
2.Wash twice at 500 g for 1 min using the washing buffer.
3. Resuspend in 0.5 mL of washing buffer and 0.5 mL of HCl 4 M.
4. Mix accurately and incubate for 30 min at room temperature.
5. Wash once as in step 2.
6. Resuspend in 1 mL of Borax buffer.
7. As in step 5.
8. Resuspend in 200 mL of washing buffer and label with 5 mL of mAb ant-BrdU.
9. Incubate for 1 hour at 4 °C in the dark.
10. As in step 5.
11. Resuspend in 200 mL of washing buffer and label with 4 mL of goat-anti-mouse FITC-conjugated antibody.
12. Incubate for 30 min at 4 °C in the dark.
13. As in step 5.
14. Resuspend in 200 mL of washing buffer and 200 mL of PI buffer.
15. Incubate for 15-30 min at 4 °C in the dark.
16. Analyse with flow cytometer equipped with a 488 nm argon laser.