Clone: 3A4Other Names: Leu-19, NKH1Isotype: Mouse IgG1, κ Description: CD56 is a single transmembrane glycoprotein also known as N-CAM(Neural Cell Adhesion Molecule),Leu-19, or NKH1. It is a member of the lg superfamily. The 140 kD isoform is expressed on NK cells and NK-T cells. CD56 is also expressed in brain (cerebellum and cortex) and at neuromuscular junctions.Certain large granular lymphocyte(LGL) leukemias, small-cell lung carcinomas, neuronal derived tumors, myelomas, and myeloid leukemias also express CD56. CD56 plays a role in hemophilic and heterophilic adhesion via binding to itself or heparin sulfate.Antibody Type: MonoclonalHost Species: MouseImmunogen: Human CD56 Recombinant ProteinFormulation: Antibodies are supplied in buffer containing stabilizer and 0.05% sodium azidePreparation: The antibody was purified by affinity chromatography, and conjugated with PE under optimal conditions.Storage & Handling: Store protected from light at 2-8°C. Do not freeze. The expiration date is indicated on the vial label.Recommended Usage: 0.25ug/TExcitation Laser: Blue Laser (488 nm) Green Laser (532 nm)/Yellow-Green Laser (561 nm) LWB Cell Preparation 1. For each patient sample, label three tubes A, B, and C. Also label each tube with the sample identification number. 2. Place 50 µL PBS reagent into Tube A, Mouse IgG1 PE Control reagent into Tube B, and 0.25ug of the CD56-PE reagent into Tube C. 3. For each sample tube, use a fresh micropipettor tip and carefully add 50 µL of human Peripheral blood into the bottom of each of the three labeled tubes. All tubes refill to 100ul. 4. Vortex thoroughly at low speed for 3 seconds and incubate for 15minutes at room temperature (20°C–25°C). Repeat this step again. NOTE Protect samples from direct light during this incubation procedure and exercise care to prevent blood from running down the side of the tube. If whole blood remains on the side of the tube, it will not be stained with the reagent. 5. Add 250uL of room temperature (20°C–25°C) lysing solution to each tube. Immediately vortex thoroughly at low speed for 3 seconds and incubate for 10– 12 minutes at room temperature (20°C–25°C) in the dark. Do not exceed 12 minutes. 6. Immediately after incubation, add 1 mL of PBS with 1% FBS to each tube. Centrifuge tubes at 1500rpm for 5 minutes at room temperature (20°C–25°C). 7. Aspirate the supernatant leaving approximately 50 µL of residual fluid in the tube to avoid disturbing the pellet. 8. Repeat 6,7steps again. 9. Vortex thoroughly at low speed to resuspend the cell pellet in the residual fluid and then add 0.3 mL of PBS to each tube. Vortex thoroughly at low speed for 3 seconds. Make sure the cells are well mixed with the fixing solution. 10. The cells are now ready to be analyzed on the flow cytometer. Cap or cover the prepared tubes and store at 2°C– 8°C in the dark until flow cytometric analysis. NOTE Analyze the fixed cells within 2 hours after staining.Over 2 hours add 0.3 mLof PBS with 1% paraformaldehyde to each tube.
