NM LYSE: Flow Cytometry Lysing Solution

For Wash- or No-Wash Lysing Procedures with Whole Blood or Marrow Samples NM-LYSE is a premixed, ready to use lysing solution fomulated for lysing erythrocytes following monoclonal antibody staining of whole blood. Treatment with this reagent simultaneously 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 populations derived from various Human biological samples (blood, bone marrow and others) using flow cytometry. 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. 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 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.

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 conjµgate - 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 conjµgate - 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 - Centrifµge 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

For professional users only. NM-LYSE contains fomaldehyde. Formaldehyde is toxic, allergenic and a suspected carcinogen. Avoid contact with eyes, skin and clothing. Proper handling procedures are recommended.

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.

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.