FITC Labeling Borrelia burgdorfori (fixed)
(created by Qilong Xu on 2008-07-27 (updated at 2008-07-27 20:14:51)
Reagents
antibody-FITC.IgG
1X PBS buffer
primary antibody (anti-DbpA serum, anti-DbpB serum, OspC mAb, FlaB mAb...)
Procedures
1. Harvest 0.6 ml of cells
2. Resuspend in 50 μl of 1x PBS buffer throughly.
3. Add 600 μl of acetone, mix with pippete tip, incubate for 15 min at room temperature.
4. Spin down for 8 min at 10,000 rpm.
5. Wash once with 800 μl 1x PBS buffer
6. Resuspend in 100 μl 1x PBS buffer
7. Add primary antibody, incubate @ 33°C for hour (tap tubes several times).
8. Wash with 500 μl of 1x PBS buffer.
9. Resuspend in 100 μl 1x PBS buffer.
10. Add secondary antiboy-FITC.IgG, incubate @33°C for hour (tap tubes several times).
11. Spin down. Discard supernatant, resuspend in 50 μl 1x PBS buffer
12. Check