COMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti CD22 (PE)

COMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti CD22 (PE)

• Background: Myeloperoxidase (MPO) is a glycoprotein present in the azurophil (primary) granules of myeloid cells, which appears in the myeloblast stage of myeloid cell differentiation. MPO is he most common functional protein of myeloid cells and is involved in the inflammatory response. It helps to kill microbes by breaking down peroxide in the presence of halide ions, contributing to the bactericidal function of granulocytes. The primary translation product of MPO undergoes glycosylation with production of the 89 kDa heme-free apopro-MPO form followed by incorporation of heme and conversion into the enzymatically active pro-MPO form. Subsequently, pro-MPO becomes targeted to azurophil granules where final processing occurs to produce mature dimeric MPO consisting of the 59-64 kDa MPO ?-chain and the 14 kDa MPO ?-chain. Precursor B-cells are surface-CD22 negative, but cytoplasmic CD22 positive. Mature B-lymphocytes.express CD22 also on their surface. The combined staining for MPO and CD22 allows the distinction of mature/immature myeloid cells and B-lymphocytes. The MPO-C2/CD22 COMBI-IC mAb permits the identification and enumeration of normal and malignant myelomonocytic cells and B lymphoid commited cells in Human blood and bone marrow 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. 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.
• CAS Number: 9007-83-4
• Host: Mouse
• Species Reactivity: Human
• Isotype: IgG1
• Clone: 8E6 and RFB4
• Conjugation: FITC, PE
• Type: Primary Antibodies
• Applications: Flow Cytometry
• Assay Principle: Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX,PERM? (Cat. No. GAS-002) intracellular MPO-C2 and CD22 can be easily stained in cell suspensions. - For each sample to be analyzed add 50 µL of whole blood, bone marrow or mononuclear cell suspension in a 5mL tube - Add 100 µL of Reagent A (Fixation Medium, stored and used at room temperature) - Incubate for 15 minutes at room temperature - Add 5mL phosphate buffered saline and centrifµge cells for 5 minutes at 300 g - Remove supernatant and add to cell pellet 100 µL Reagent B (Permeabilization Medium) and 20 µL of the MPO-C2/CD22 COMBI-IC monoclonal antibody conjµgate - Vortex at low speed for 1-2 seconds - Incubate for 15 minutes at room temperature - Wash cells with phosphate buffered saline as described above - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 mL 1.0 % formaldehyde and store them at 2-8°C in the dark. Analyze fixed cells within 24 hours.
• Form: 1 mL of FITC-conjµgated anti Myeloperoxidase-C2 (clone 8E6) and PE-conjµgated anti CD22 (cloneRFB4) in PBS pH 7,2, 1% BSA, and 0,05% NaN3, approximately 50 tests
• Precautions: For professional users only. This reagent contains sodium azide. To avoid the development of hazardous conditions, reagents containing azide should be diluted in running water prior to be discarded. Similar to the work with other biological products, proper handling procedures are recommended.
• References & Citations: 1. Andersson, E., Hellman, L., Gullberg, U. , Olsson, I. (1998) J Biol Chem 273, 4747-53. _x000D_ 2. Beverley, P. C., Linch, D. , Callard, R. E. (1981) Haematol Blood Transfus 26, 309-13. _x000D_ 3. Boue, D. R. , Lebien, T. W. (1988) J Immunol 140, 192-9. _x000D_ 4. Braylan, R. C., Orfao, A., Borowitz, M. J. , Davis, B. H. (2001) Cytometry 46, 23-7_x000D_ 5. Catovsky, D., Matutes, E., Buccheri, V., Shetty, V., Hanslip, J., Yoshida, N. , Morilla, R. (1991) Ann Hematol 62, 16-21._x000D_ 6. Janossy, G., Coustan-Smith, E. , Campana, D. (1989) Leukemia 3, 170-81. _x000D_ 7. Knapp, W., Majdic, O. , Strobl, H. (1993) Recent Results Cancer Res 131, 31-40. _x000D_ 8. Koeffler, H. P., Ranyard, J. , Pertcheck, M. (1985) Blood 65, 484-91. _x000D_ 9. Konikova, E., Glasova, M., Kusenda, J. , Babusikova, O. (1998) Neoplasma 45, 282-91. _x000D_ 10. Lanza, F., Latorraca, A., Moretti, S., Castagnari, B., Ferrari, L. , Castoldi, G. (1997) Cytometry 30, 134-44. _x000D_ 11. Li, J. L., Shen, G. L., Ghetie, M. A., May, R. D., Till, M., Ghetie, V., Uhr, J. W., Janossy, G., Thorpe, P. E., Amlot, P. , et al. (1989) Cell Immunol 118, 85-99. _x000D_ 12. Mason, D. Y., Stein, H., Gerdes, J., Pulford, K. A., Ralfkiaer, E., Falini, B., Erber, W. N., Micklem, K. , Gatter, K. C. (1987) Blood 69, 836-40. _x000D_ 13. Murao, S., Stevens, F. J., Ito, A. , Huberman, E. (1988) Proc Natl Acad Sci U S A 85, 1232-6.
• Storage Conditions: Nordic-MUbio monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody; For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8°C (DO NOT FREEZE!) and protected from prolonged exposure to light. If a slight precipitation occurs upon storage, this should be removed by centrifugation; It will not affect the performance or the concentration of the product 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.