CD45 Antibody – Mouse Monoclonal

Monoclonal

Mouse IgG1

OX-1

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 ascitic fluid via Protein G Chromatography))

Antibody Concentration: 0.1 mg/ml (lot specific)

CD45 (LCA) (Rat Leukocyte Common Antigen)

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.

chromosome: Un

Rat thymocyte membrane glycoproteins

BALB/c Spleen

NSI/1-Ag4-1

100 ug or 500 ug 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.

1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population concentration to 4-5 mg/ml with Lympholyte-M 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 1x10⁶ cells, representing 1 test).
4. To each tube, add 1.0-0.5 ug of MBS520138 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-FITC) at a 1:500 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. Was 2 times at 4°C
11. Resuspend the cell pellet in 50 ul ice cold media B.
12. Transfer to suitable tubes for flow cytometric analysis contianing 15 ul of propidium iodide at 0.5 mg/ml in PBS. This satins dead cells by intercalating in DNA.

A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 ul of 2 M sodium azide in 100 mls.)
B. Phosphate buffered saline (pH 7.2) + 5% bovine serum albumin + sodium azide (100 ul of 2 M sodium azide in 100 mls.)

Procedure: As above
Antibody Concentration: 0.05ug/10⁶ cells
Strains Tested: Wistar, Fischer 344
Positive: Wistar, Fischer 344
Negative: None