Flow cytometry (see protocol below). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protokoll
FLOW CYTOMETRY ANALYSIS: 1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood cellpopulation. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube add 0. 2- 0. 5 mg of antibody per 10e6 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. Resuspend the cell pellet in 50 µl ice cold Media B. 9. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in phosphate buffered saline. (This stains dead cells by intercalatingDNA. )MEDIA:
Beschränkungen
Nur für Forschungszwecke einsetzbar
Konzentration
0.1 mg/mL
Buffer
PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL.
Konservierungsmittel
Sodium azide
Vorsichtsmaßnahmen
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung
Avoid repeated freezing and thawing.
Lagerung
-20 °C
Informationen zur Lagerung
Prior to reconstitution store at -70 °C. Following reconstitution store the antibody (in aliquots) at -20 °C for 6 month.