CD25 Antibody – Mouse Monoclonal

Monoclonal

Mouse IgG1

MRC OX-39

Mouse

Rat

Biotin (Biotin conjugated Ig buffered in PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. (Purified from bioreactor supernatant))

0.1 mg/ml (lot specific)

CD25 (IL-2Ra) (Rat CD25 (IL-2R))

Store at 4°C. For long term storage, aliquot and freeze unused portion at -20°C in volumes appropriate for single usage.

Avoid freeze/thaw cycles.

ModBase 3D Structure for P25094

Immunization:

Immunogen:
T blasts from a mixed lymphocyte reaction between purified CD4 positive T cells and irradiated spleens.

BALB/c spleen

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 ?l of this suspension to each tube (each tube will then contain 1 x 106 cells, representing 1 test).
4. To each tube, add 1.0-.5ug* of MBS520072 per 106 cells.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 (Streptavidin-PE) at a 1:50 dilution.
9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
10. Wash 2 times at 4°C.
11. Resuspend the cell pellet in 50 ?l ice cold media B.
12. Transfer to suitable tubes for flow cytometric analysis containing 15 ?l of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.

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 (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 ul of 2M sodium azide in 100 mls).