Description

Description: 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 determination of cellular subpopulations in non-separated biological samples. This is not only time saving but reduces also the probability of an unintended loss of distinct cellular populations due to e.g. commonly used differential centrifugation procedures. With the NM-LYSE reagent flow cytometric analysis of whole blood has become as easy and accurate as the analysis of separated cell populations. Results must be interpreted by a certified professional before final interpretation. Analyses performed with this antibody should be paralleled by positive and negative controls. If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us.

Additional Text

Antibody Clonality

Monoclonal

Application Notes

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 4°C 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 4°C 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.

Storage Note

NM-LYSE reagent should be stored and used at room temperature. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended. Do not use reagent if a precipitate should form or discoloration occurs. If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us