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Category
Other Reagents
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Long description
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.
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Antibody come from
n/a
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Other description
No other information available
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Clone
not specified
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Antigen antibody binding interaction
NM LYSE: Flow Cytometry Lysing Solution
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Antibody is raised in
see techfile
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Antibody s reacts with
see techfile
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Antibody s reacts with these species
This antibody doesn't cross react with other species
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Antibody s specificity
NM-LYSE Reagent is designed for use with all commercially available flow cytometers. Alignment and compensation should be performed according to manufacturer´s instructions 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.
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Application
Flow Cytometry
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Antibody s suited for
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
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Storage
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
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Relevant references
Bossuyt, X., Marti, G. E. & Fleisher, T. A. (1997) Cytometry 30, 124-33. Fritsch, G., Printz, D., Stimpfl, M., Dworzak, M. N., Witt, V., Potschger, U. & Buchinger, P. (1997) Transfusion 37, 775-84. Kormoczi, G. F., Wolfel, U. M., Rosenkranz, A. R., Horl, W. H., Oberbauer, R. & Zlabinger, G. J. (2001) J Immunol 167, 451-60. 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.
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Protein number
see ncbi
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Warnings
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.
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Description
FACS or Flow Cytometry is an SSC and FSC analysis by scattered light gating. Becton, BD or coulter facses analyze for the major part human CD marker antibodies and cellular markers by PE or FITC labelled antibodies. Facses or flow cytometers will analyze forward and side scatters by gating of human lymphocytes. Becton Dickinson uses anti human CD antigens, CD4, CD8 monoclonals. The clones of these antibodies have a known affinity to these membrane receptors. The volume (ml) in milliliters of buffered % w/v solutions at the medium pH are also used for DNA extraction Organic (Phenol-Chloroform) Extraction, Non-Organic (Proteinase K and Salting out), Chelex (Ion Exchange Resin) Extraction, EDTA or PBS aqueous.
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Test
Flow cytometry uses monoclonal antibodies of specific affinity clones for cell counting, cell sorting and biomarker detection by suspending cells in a stream of fluid for Forward Scatter, FSC and side scatter, SSC analysis. Human PBMCs can be loaded with CFSE tracking dye after non adherent cell harvesting. Subsequently labeled with anti-CD antibodies, and analyzed by multiparameter flow cytometry. Two-parameter profiles of CD vs. CFSE; and another CD vs. FSC-W. We suggest to use FSC-H vs. FSC-A. FSC-A, FSC-H, FSC-W = area, height, and width of the forward 488 nm light scatter from the flow signal.
