gamma/delta TCR Antibody – Mouse Monoclonal

Flow Cytometry Analysis:
Method:
1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-Rat cell separation medium.
2. Wash 2 times.
3. Resuspend the cells to a concentration of 2x10⁷ cells/ml in media A. Add 50 ul of this suspension to each tube (each tube will then contain 1 x 10⁶ cells, representing 1 test).
4. To each tube, add 0.5- 1.0 ug* of MBS520299 .
5. Vortex the tubes to ensure thorough mixing of antibody and cells.
6. Incubate the tubes for 30 minutes at 4°C.
7. Wash 2 times at 4°C.
8. Add 100 ul of secondary antibody (PE-Goat anti-mouse IgG (H+L)) at 1:20 dilution.
9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive).
10. Wash 2 times at 4°C in media B.
11. Resuspend the cell pellet in 50 ul ice cold media B.
12. Transfer to suitable tubes for flow cytometric analysis containing 15 ul of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.

Media:
A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 ul of 2M sodium azide in 100 mls).
B. Phosphate buffered saline (pi I 7.2) + 0.5% Bovine serum albumin + sodium azide (100 ul of 2M sodium azide in 100 mls).