Clone: 4A5Other Names: FcγRIIIIsotype: Mouse IgG2b, κ Description: CD16 is a low affinity Fc receptor.It is a cluster of differentiation molecule found on the surface of natural killer cells, neutrophil polymorphonuclear leukocytes, monocytes and macrophages. It can be used to isolate populations of these cells by antibodies directed towards CD16, using fluorescent-activated cell sorting or magnetic-activated cell sorting.CD16 has been identified as Fc receptors FcγRIIIa (CD16a) and FcγRIIIb (CD16b). These receptors bind to the Fc portion of IgG antibodies which then activates the NK cell for antibody-dependent cell-mediated cytotoxicity. A lack of CD16 in a given population of neutrophils may indicate prematurity, as could be caused by a left shift due to neutrophilic leukocytosis induced by tissue necrosis or bacterial infection.Antibody Type: MonoclonalHost Species: MouseImmunogen: Human CD16 Recombinant ProteinFormulation: Antibodies are supplied in buffer containing stabilizer and 0.05% sodium azidePreparation: The antibody was purified by affinity chromatography.Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.Recommended Usage: 0.5ug/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. Label each tube with the sample identification number. 2. Place 50 µL PBS reagent into Tube A, Mouse IgG2b Control reagent into Tube B, and 0.5µg of the CD16 reagent into Tube C. 3. For each sample tube, use a fresh micropipettor tip and carefully add 5*105 pbmc 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 4℃. Then repeat this step again. NOTE Protect samples from direct light during this incubation procedure . 5. 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). 6. Aspirate the supernatant leaving approximately 50 µL of residual fluid in the tube to avoid disturbing the pellet. 7. Repeat 5,6 steps twice. 8. The secondary antibody used was Beckman Goat F（ab')2 Fragment Anti mouse IgG(H+L)-PE (BECKMAN COURTER IM0855) secondary antibody at 1/1000 dilution. 9. Place 100 µL secondary antibody into Tube B,C. Vortex thoroughly at low speed for 3 seconds and incubate for 15minutes at 4℃. Then repeat this step again. 10. Repeat 5,6 steps twice. 11. Add 0.3 mL of PBS to each tube and then vrtex thoroughly at low speed to resuspend the cell pellet in the residual fluid . Vortex thoroughly at low speed for 3 seconds. Make sure the cells are well mixed with the fixing solution. 12. 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. For research use only. Not for diagnostic use. Not for resale. Suzhou Bright Scistar Biotech Co. Ltd will not be held responsible for patent infringement or other violations that may occur with the use of our products.
