Clone: LZ-2 and 4C5
Label: FITC/PE
Background: Lysozyme (LZ) is a cationic antimicrobial peptide of 14 kDa. Lysozyme is stored in primary but predominantly in specific (secondary) granules of neutrophils. It cleaves peptidoglycan constituents of the bacterial cell wall and can bind LPS. The epitope recognized by antibody LZ-2 is expressed by virtually all myeloid cells including normal and malignant granulocytes and monocytes. In normal myelopoiesis LZ can first be detected at the myeloblast stage where it appears somewhat later than MPO expression. Lactoferrin (LF) is an iron-binding protein with bactericidal and bacteriostatic activity, which is stored within the secondary granules of granulocytes. LF expression is restricted to the post-mitotic maturation compartment of the granulocytic lineage, starting from the myelocyte stage. Normal and malignant myeloblasts are LF negative. The combined staining for Lysozyme and Lactoferrin allows the distinction between mature and immature myelomonocytic cells. The LZ/LF COMBI-IC reagent permits the identification and enumeration of myeloid cells in normal and malignant human blood and bone marrow samples 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
Specificity: The anti-Lysozyme Antibody (clone LZ-2) reacts with intracellular human lysozyme/muramidase expressed by virtually all myelomonocytic cells, macrophages and their precursors. The LF mAb (clone 4C5) recognizes lactoferrin stored within secondary granules of postmitotic granulocyte-commited cells.In this COMBI-IC Reagent antibody LZ-2 is conjugated to FITC, antibody 4C5 is conjugated to Phycoeythrin (PE).The sensitivity y of LZ/LF mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAbdilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI (mean fluorescence intensity). For this purpose, a mAb-concentration range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilutionexpected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50 µl of leukocytes containing 10^7 cells/ml are stained with 20µl mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection threshold. The final concentration of the product is then adjusted to be at least 3-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.
Formulation: PBS pH 7.2, 1% BSA, 0.05% NaN3
References: 1. Knapp, W., Majdic, O. & Strobl, H. (1993) Recent Results Cancer Res 131, 31-40. 2. Konikova, E., Glasova, M., Kusenda, J. & Babusikova, O. (1998) Neoplasma 45, 282-91. 3. Lanza, F., Latorraca, A., Moretti, S., Castagnari, B., Ferrari, L. & Castoldi, G. (1997) Cytometry 30, 134-44. 4. Oehler, L., Majdic, O., Pickl, W. F., Stockl, J., Riedl, E., Drach, J., Rappersberger, K., Geissler, K. & Knapp, W. (1998) J Exp Med 187, 1019-28. 5. Paietta, E. (2003) Best Pract Res Clin Haematol 16, 671-83. 6. Rado, T. A., Bollekens, J., St Laurent, G., Parker, L. & Benz, E. J., Jr. (1984) Blood 64, 1103-9. 7. Srivastava, C. H., Rado, T. A., Bauerle, D. & Broxmeyer, H. E. (1991) J Immunol 146, 1014-9. 8. Strobl, H. & Knapp, W. (2004) J Biol Regul Homeost Agents 18, 335-9. 9. Teng, C. T., Gladwell, W., Beard, C., Walmer, D., Teng, C. S. & Brenner, R. (2002) Mol Hum Reprod 8, 58-67. 10. Braylan, R. C., Orfao, A., Borowitz, M. J. & Davis, B. H. (2001)Cytometry 46, 23-7. 11. Catovsky, D., Matutes, E., Buccheri, V., Shetty, V., Hanslip, J., Yoshida, N. & Morilla, R. (1991) Ann Hematol 62, 16-21.
Caution: 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.
English
Deutsch
