Cytotoxicity assay (see protocol). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protokoll
RECOMMENDED METHOD FOR DEPLETING A CELL POPULATION OF H-2Kb AND/OR H-2Db BEARING LYMPHOCYTES: 1. Prepare a cell suspension from the appropriate tissue in Cytotoxicity Medium1 orequivalent. Remove red cells and dead cells (where necessary) by purification of viablelymphocytes on density cell separation medium. After washing, adjust the cellconcentration to 1x10e7 cells per ml in Cytotoxicity Medium. 2. Add the antibody to a final concentration of 1: 100 and mix. Alternatively, pellet the cellsand resuspend in antibody diluted 1: 100 in Cytotoxicity Medium. 3. Incubate for 60 minutes at 4°C. 4. Centrifuge to pellet the cells and discard the supernatant. 5. Resuspend to the original volume in Rabbit Complement, diluted to the appropriateconcentration in Cytotoxicity Medium. (Recommended concentration included with eachbatch of Rabbit Complement. )6. Incubate for 60 minutes at 37°C. 7. Monitor for percent cytotoxicity at this stage, before further processing. For this purposeremove a small sample from each tube, dilute 1: 10 with medium, and add 1/10 volumeof 1% Trypan Blue. After 3-5 minutes, score live versus dead cells in a haemacytometer.
Beschränkungen
Nur für Forschungszwecke einsetzbar
Konzentration
1.0 mg/mL
Buffer
PBS + 0.02 % NaN3
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. Dilute only prior to immediate use
Lagerung
4 °C/-20 °C
Informationen zur Lagerung
Store the antibody (undiluted) at 2-8 °C for one month or (in aliquots) at -20 °C for longer.