CD16 antibody (FITC)

Catalog No. ABIN1112067

Product Details anti-CD16 Antibody (FITC)

Target: CD16

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD16 antibody is conjugated to FITC

Application: Flow Cytometry (FACS), Immunofluorescence (IF)

Characteristics: Monoclonal Mouse Anti-Human CD16 FITC, is recommended for use in flow cytometry for identification antigen (Fc RII) present on NK cells, neutrophils and basophils in peripheral blood and bone marrow.

Clone: GRM1

Isotype: IgG1

Application Details

Application Notes: It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence staining of human tissue for flow cytometric analysis using 20 µl/10^6 cells.

Comment:

Fluorescein isothiocyanate (Molecular Probes).

Sample Collection: 1. Transfer 100 µl of anticoagulated (EDTA) blood to a 12 x 75 mm polystyrene test tube (10^6 cells). 2. Add 20 µl of CD16 FITC and mix gently with a vortex mixer. The 20 µl is a guideline only, the optimal volume should be determined by the individual laboratory. 3. The recommended negative control is a non-reactive FITC-conjugated antibody of the same isotype. 4. Incubate in the dark at room temperature at 4°C for 30 minutes or at room temperature (20-25 °C) for 15 minutes. 5. Add 1,5 ml of Lysing Solution to each sample and mix gently with a vortex mixer. Incubate for 10 minutes at room temperature in the dark. 6. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 7. Add 2 ml 0.01 mol/l PBS (It betters that it containing 2% bovine serum albumin) and resuspend the cells by using a vortex mixer. 8. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 9. Resuspend pellet in an appropriate fluid for flow cytometry, e.g. 0.3 ml PBS. The PBS should contain 1% paraformaldehyde (fixative) if samples are not analysed the same day. 10. Analyse on a flow cytometer or store at 2-8 °C in the dark until analysis. Samples can be run up to 24 hours after lysis.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: The conjugate is provided in liquid form in buffer containing 1% bovine serum albumin (BSA) and 0,09% Sodium azide, pH 7.2.

Preservative: Sodium azide

Precaution of Use: 1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it must not be used in conjunction with patient records or treatment. 2. This product contains Sodium azide (NaN3), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, Sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used.

Storage: 4 °C

Target Details for CD16

Target: CD16

Alternative Name: CD16 (CD16 Products)

Synonyms: Fcgr3 antibody, Fcgr3a antibody, CD16 antibody, FCGR3 antibody, FCGR3@ antibody, CD16b antibody, FCG3 antibody, FCR-10 antibody, FCRIII antibody, FCRIIIb antibody, FcRIII antibody, Fc fragment of IgG, low affinity IIa, receptor antibody, Fc fragment of IgG, low affinity IIIa, receptor (CD16a) antibody, Fc fragment of IgG receptor IIIb antibody, Fc fragment of IgG receptor IIIa antibody, Fc receptor, IgG, low affinity III antibody, Fc-gamma RIII antibody, Fcgr2a antibody, FCGR3A antibody, FCGR3B antibody, Fcgr3 antibody, FCGR3 antibody

Background: The CD16 molecule has been described as the low affinity Fc receptor (FcRII) for complexed IgG which may exist either as a transmembranous form or as glycosyl phosphatidylinositol form. It is expressed on NK cells, granulocytes (PMN) and macrophages. The GPI-linked form is also expressed on neutrophils. We have produced a monoclonal antibody, GRM1, against a prolymphocytic leukemia that defines an antigen present in neutrophilic granulocytes (PMN) and a lymphocyte subset with natural killer (NK) activity, which was identified as large granular lymphocytes. This monoclonal antibody recognizes FcR2 (CD16), an antigen composed of two polypeptides of 50 and 60 kDa, respectively. This GRM1 monoclonal antibody was tested against normal T and B cells, neutrophilic granulocytes, monocytes, platelets, acute and chronic leukemias, and was positive only against granulocytes (95%) and cells with NK activity. GRM1 was able to deplete NK cell activity in complement-dependent lysis. However, GRM1 did not block NK activity nor peripheral blood lymphocyte- and PMN-mediated antibody-dependent cytotoxicity in healthy individuals. GRM1 also did not block Fc receptor in an erythrocyte antibody rosette assay. The immunochemical data and cell distribution patterns lead us to conclude that GRM1 recognizes and FcR2 receptor epitope which is not involved in the receptor's function.

Pathways: Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process