Mouse anti Lysozyme conugated to FITC

Permeabilization and Staining Procedure
- In combination with our Permeabilization Kit FIX&PERM? (Cat. No. GAS-002) intracellular Lysozyme can be easily stained in cell suspensions.
- For each sample to be analyzed add 50 ?l of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube
- Add 100 ?l 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 ?l Reagent B (Permeabilization Medium) and 20 ?l of the LZ-2 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.

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.

The LZ-2 antibody permits the identification and enumeration of human myelomonocytic cells 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.

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.

Purified by Chromatography, Storage buffer: PBS pH 7.2, 1% BSA, 0.05% NaN3

FITC

2 ml of FITC-conjugated anti Lysozyme (clone LZ-2) in PBS pH 7.2, 1% BSA, and 0.05% NaN3, approximately 100 tests.

FITC

Purified by Chromatography

The anti-Lysozyme Antibody (clone LZ-2) reacts with intracellular human lysozyme/muramidase expressed by virtually all myelomonocytic cells, macrophages and their precursors.

The sensitivity of LZ-2 mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAb dilutions 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 dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells).In practice, 50 ?l of leukocytes containing 10^7 cells/ml are stained with 20 ?l 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.

P61626