NM-LYSE: Flow Cytometry Lysing Solution (CE/IVD)

NM-LYSE: Flow Cytometry Lysing Solution (CE/IVD)

• Background: 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._x000D_ Results must be put within the context of other diagnostic tests as well as the clinical history of the patient 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.
• Type: Buffers and Reagents
• Applications: Flow Cytometry
• Assay Principle: 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._x000D_ _x000D_ No-wash staining and lysing procedure_x000D_ • For each sample add 50 µL of EDTA anti-coagulated blood to a 3-5 mL tube_x000D_ • Add 20 µL of the appropriate Nordic-MUbio monoclonal antibody conjµgate_x000D_ • Incubate the tube for 15 minutes at 4°C or at room temperature in the dark_x000D_ • Add 100 µL NM-LYSE to each tube and incubate for 10 minutes at room temperature_x000D_ • Add 1 mL of destilled water and vortex, incubate for 5-10 minutes at room temperature_x000D_ • Analyze immediately or store samples at 2-8° C in the dark and analyze within 24 hours_x000D_ _x000D_ Wash staining and lysing procedure_x000D_ • For each sample add 50 µL of EDTA anti-coagulated blood to a 3-5 mL tube_x000D_ • Add 20 µL of the appropriate Nordic-MUbio monoclonal antibody conjµgate_x000D_ • Incubate the tube for 15 minutes at 4°C or at room temperature in the dark_x000D_ • Add 100 µL NM-LYSE to each tube and incubate for 10 minutes at room temperature_x000D_ • Add 3-4 mL of destilled water and vortex, incubate for 5-10 minutes at room temperature_x000D_ • Centrifµge tube for 5 minutes at 300 g_x000D_ • Aspirate supernatant and resuspend pellet in 0.3 mL of sheath fluid_x000D_ • Analyze immediately or store samples at 2-8° C in the dark and analyze within 24 hours_x000D_ _x000D_ NM-LYSE is designed for use with all commercially available flow cytometers. Alignment and compensation should be performed according to manufacturer´s instructions.
• Form: 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 fomµLated for lysing erythrocytes following monoclonal antibody staining of whole blood; Treatment with this reagent simµLtaneously leads to lysis of red blood cells and fixation of white cells. Morphological scatter characteristics of leukocytes remain intact. NM-LYSE can be used with or without sample washing. NM-LYSE is suitable for the analysis of; Normal and malignant leukocyte popµLations derived from various Human biological samples (blood, bone marrow and others) using flow cytometry. ResµLts must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation._x000D_ The quality of each NM-LYSE Lot is determined by lysing red blood cells of well defined blood samples from representative donors and subsequent comparison of forward and side scatter characteristics of obtained leukocytes.
• Precautions: For professional users only. NM-LYS contains fomaldehyde and is labelled: Harmful. Formaldehyde is toxic, allergenic and a suspected carcinogen. Never pipette by mouth and avoid contact with eyes, skin and clothing. Proper handling procedures are recommended. As a main rule, persons under 18 years of age are not allowed to work with this product. Users must be carefully instructed in the proper working procedure, the dangerous properties of the product and the necessary safety instructions. Please refer to the Material Safety Data Sheet (MSDS) for additional information._x000D_ Dispose product remainders according to local regulations._x000D_ _x000D_ For research use only. Not for diagnostic or therapeutic use
• References & Citations: 1. Bossuyt, X., Marti, G. E. , Fleisher, T. A. (1997) Cytometry 30, 124-33. _x000D_ 2. Fritsch, G., Printz, D., Stimpfl, M., Dworzak, M. N., Witt, V., Potschger, U. , Buchinger, P. (1997) Transfusion 37, 775-84. _x000D_ 3. Kormoczi, G. F., Wolfel, U. M., Rosenkranz, A. R., Horl, W. H., Oberbauer, R. , Zlabinger, G. J. (2001) J Immunol 167, 451-60. _x000D_ 4. Menendez, P., Redondo, O., Rodriguez, A., Lopez-Berges, M. C., Ercilla, G., Lopez, A., Duran, A., Almeida, J., Perez-Simon, J. A., San Miguel, J. F., Gratama, J. W. , Orfao, A. (1998) Cytometry 34, 264-71.
• Storage Conditions: 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