COMBI IC Reagent: Mouse anti CD45 (FITC) and Mouse anti CD14 (PE)

Nordic-MUbio

COMBI IC Reagent: Mouse anti CD45 (FITC) and Mouse anti CD14 (PE)

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The CD45 mAb (clone VIT200) recognizes a pan CD45 epitope. The CD14 mAb (clone MEM18) recognizes surface CD14 on human monocytes and macrophages as well as neutrophils. The sensitivity of CD45/CD14 mAb is determined by staining well-defined blood samples

Staining Procedure Direct Immunofluorescence (Staining Procedure) Nordic-MUbio fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparations. Proposed staining procedure for whole blood in short: - For each sample add 50 microl of EDTA anti-coagulated blood to a 3-5 ml tube - Add 20 microl of the appropriate Nordic-MUbio monoclonal antibody conjugate - Incubate the tube for 15 minutes at 4°C or at room temperature in the dark - Add 100 microl NM-LYSE (Cat.No. GAS-003) to each tube and incubate for 10 minutes at room temperature - Add 3-4 ml of destilled water and vortex, incubate for 5-10 minutes at room temperature - Centrifuge tube for 5 minutes at 300 g - Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid - Analyze immediately or store samples at 2-8° C in the dark and analyze within 24 hours For No-Wash protocol please refer to www.nordicmubio.com Proposed staining procedure for MNC in short: - Carefully add 20 microl antibody conjugate and 50-100 microl MNC to the bottom of a tube - Vortex at low speed for 1-2 seconds - Incubate for 15-30 minutes at 2-8°C or at room temperature - Centrifuge tubes for 5 minutes at 300 g - Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge cells again for 5 minutes at 300 g - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1 % formaldehyde and store them at 2-8°C in the dark. Analyze fixed cells within 24 hours

Flow Cytometry, ImmunoFluorescence

Direct Immunofluorescence;Flow Cytometry

Primary antibodies

Research Use Only

Monoclonal

VIT200 and MEM18

FITC, RPE

Mouse

IgG1, IgG2a

Human

Human

2°C to 8°C

12352203