FIX&PERM(r) Solution A (Fix) (CE/IVD)

Nordic-MUbio

FIX&PERM Solution A (Fix) (CE/IVD)

7

Biological fluids (blood, bone marrow, and others) must be collected under sterile conditions. Anticoagulation with EDTA or heparin is recommended. The samples should be stored at room temperature until used. For optimal results, samples should be processed and analyzed within 24 hours. Samples with high numbers of non-viable cells might cause false results, such cases require determination of cell viability with e.g. propidium iodide. All biological samples have to be handled with caution. Always consider them as potentially infective. Use appropriate precautions such as gloves, lab-coat, etc. Fixation, permeabilization and staining procedure (when combined with Reagent B / GAS-002B): For each sample to be analyzed add 50 microl of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube Add 100 microl 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 microl Reagent B (Permeabilization Medium) and 20 microl of the appropriate Nordic-MUbio 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. Comments: Special cases (diluted bone marrow samples, other samples containing low soluble protein) might benefit from replenishment with plasma components before the FIX&PERM® treatment in order to create a milieu, which more closely resembles the stuation in anti-coagulated blood. For that pupose addition of IgG preparations (e.g. Beriglobulin P, ZLB Behring, final concentration 10mg/ml) and human serum albumin (e.g. human albumin Behring 20% - infusion solution, final concentration 40mg/ml) is recommended.

Flow Cytometry

Buffers and reagents

Research Use Only

RT

41116124