Description

Description: 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 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.

Additional Text

Antibody Clonality

Monoclonal

Application Notes

Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM? (Cat. No. GAS-002) intracellular Lysozyme and Lactoferrin 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 centrifuge 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 LZ/LF COMBI-IC monoclonal antibody conjugate - 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.

Storage Note

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.