Catalog# / Size : XG-AB2005（25T/50T/100T）
Description
Species Reactivity | Human |
Clone | 4A5 |
Other Names | FcγRIII |
Isotype | 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. |
Product Details
Antibody Type | Monoclonal |
Host Species | Mouse |
Immunogen | Human CD16 Recombinant Protein |
Formulation | Antibodies are supplied in buffer containing stabilizer and 0.05% sodium azide |
Preparation | The antibody was purified by affinity chromatography, and conjugated with FITC 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. |
Application | FC - Quality tested |
Recommended Usage | 0.25ug/T |
Excitation Laser | Blue Laser (488 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 IgG2b FITC Control reagent into Tube B, and 0.25ug of the CD16-FITCreagent 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. |
Data
Flow Cytometry | |
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Human peripheral blood lymphocytes stained with CD16(4A5) PE | |
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