Flow cytometric analyses with monoclonal antibodies were so far restricted to leukocyte populations, which had been separated from erythrocytes before staining and/or analysis. Instead, whole blood staining methods allow for a rapid and accurate determin
Puffer:
NM-LYSE can be applied in wash- or no-wash lysing procedures with whole blood or bone marrow samples. NM-LYSE is a premixed, ready to use lysing solution fomulated for lysing erythrocytes following monoclonal antibody staining of whole blood. Treatment wi
Anwendungsbeschreibung:
Biological fluids (blood, bone marrow, and others) must be collected under sterile conditions. Anticoagulation with EDTA or heparin is recommended. The samples should be stored at room temperature until used. For optimal results, samples should be processed and analyzed within 24 hours. Samples with high numbers of non-viable cells might cause false results, such cases require determination of cell viability with e.g. propidium iodide. All biological samples have to be handled with caution. Always consider them as potentially infective. Use appropriate precautions such as gloves, lab-coat, etc. No-wash staining and lysing procedure For each sample add 50 µl of EDTA anti-coagulated blood to a 3-5 ml tube Add 20 µl of the appropriate Nordic-MUbio monoclonal antibody conjugate Incubate the tube for 15 minutes at 4C or at room temperature in the dark Add 100 µl NM-LYSE to each tube and incubate for 10 minutes at room temperature Add 1 ml of destilled water and vortex, incubate for 5-10 minutes at room temperature Analyze immediately or store samples at 2-8 C in the dark and analyze within 24 hours Wash staining and lysing procedure For each sample add 50 µl of EDTA anti-coagulated blood to a 3-5 ml tube Add 20 µl of the appropriate Nordic-MUbio monoclonal antibody conjugate Incubate the tube for 15 minutes at 4C or at room temperature in the dark Add 100 µl NM-LYSE to each tube and incubate for 10 minutes at room temperature Add 3-4 ml of destilled water and vortex, incubate for 5-10 minutes at room temperature Centrifuge tube for 5 minutes at 300 g Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid Analyze immediately or store samples at 2-8 C in the dark and analyze within 24 hours NM-LYSE is designed for use with all commercially available flow cytometers. Alignment and compensation should be performed according to manufacturer´s instructions.
GAS-003
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