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 ul of leukocytes containing 10^7 cells/ml are stained with 20ul 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.
