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Home > Antibodies > Anti-Human Antibodies (In Vivo) > Anti-Human CD16 Monoclonal Antibodies

Anti-Human CD16 Monoclonal Antibodies

cluster of differentiation 16, Fc gamma RIIIA and Fc gamma RIIIB, Human Fc Receptor Blocking Antibody

Catalog No. Product Name Size List Price (US$) Quantity
PA007277.m1 In vivo Grade Recombinant Anti-human CD16 Monoclonal Antibody (Clone: 3G8), Mouse IgG1 Kappa 1 mg 150.00
PA007277.m1 In vivo Grade Recombinant Anti-human CD16 Monoclonal Antibody (Clone: 3G8), Mouse IgG1 Kappa 5 mg 350.00
PA007277.m1 In vivo Grade Recombinant Anti-human CD16 Monoclonal Antibody (Clone: 3G8), Mouse IgG1 Kappa 25 mg 900.00
PA007277.h1Fs In vivo Grade Recombinant Anti-human CD16 Monoclonal Antibody (Clone: 3G8), Human IgG1 Fc Silent Kappa 1 mg 150.00
PA007277.h1Fs In vivo Grade Recombinant Anti-human CD16 Monoclonal Antibody (Clone: 3G8), Human IgG1 Fc Silent Kappa 5 mg 350.00
PA007277.h1Fs In vivo Grade Recombinant Anti-human CD16 Monoclonal Antibody (Clone: 3G8), Human IgG1 Fc Silent Kappa 25 mg 900.00
Description

PA007277.m1: In vivo Grade Recombinant Anti-human CD16 Monoclonal Antibody, Mouse IgG1 Kappa (Clone: 3G8)

Recombinant mouse IgG1 Monoclonal Antibody. 
Clone: 3G8.
Isotype: Mouse IgG1 kappa.
Source: The anti-human CD16 monoclonal antibody (clone: 3G8) was produced in mammalian cells.
Specificity/Sensitivity: The in vivo grade recombinant mouse monoclonal antibody (clone: 3G8) specifically binds to human and non-human primate CD16.
Applications: ELISA, flow cytometry, neutralization, functional assays such as bioanalytical PK and ADA assays, and those assays for studying biological pathways affected by the human CD16 protein. Human Fc receptor blocking solution, human Fc receptor blocking reagent, and human Fc receptor blocking antibody.
Form of Antibody: 0.2 uM filtered solution, pH 7.4, no stabilizers or preservatives.
Endotoxin: < 1 EU per 1 mg of the protein by the LAL method.
Purity: >95% by SDS-PAGE under reducing conditions and HPLC.

Shipping: The antibody is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70°C as supplied.
1 month from date of receipt, 2 to 8°C as supplied.

Background

The recombinant 3G8 antibody binds to human and non-human primate CD16 or FcγRIII, a cluster of differentiation molecule found on the surface of natural killer cells, neutrophils, monocytes, macrophages, and certain T cells. CD16 consists of Fc receptors FcγRIIIa (CD16a) and FcγRIIIb (CD16b). The most well-researched membrane receptor implicated in triggering lysis by NK cells, CD16 is a molecule of the immunoglobulin superfamily (IgSF) involved in antibody-dependent cellular cytotoxicity (ADCC). It can be used to isolate populations of specific immune cells through fluorescent-activated cell sorting (FACS) or magnetic-activated cell sorting, using antibodies directed towards CD16.

The most popular human Fc blocking reagents include:
Anti-human CD16 (clone 3G8), and CD32 (Clone IV.3): for flow cytometry, immunohistochemistry (IHC), and immunoprecipitation (IP). It is highly specific for FcγRII (CD32) and FcγRIII (CD16), and reduces background staining; it may not block all Fc receptors. Clones 10.1, and 22 or H22 are the most widely used anti-human FcγRI (CD64) clone for Fc blocking in flow cytometry and functional assays. It prevents non-specific Fc receptor binding on monocytes, macrophages, and dendritic cells, improving antibody specificity.
Purified human IgGs or mixes of human isotype controls: for general Fc blocking in IHC and immunofluorescence (IF). It is cheap and easy to use but it is less specific, and may even introduce unwanted immunoglobulins. Syd Labs supply various human IgG isotype controls.
Normal mouse serum, purified mouse IgG or isotype controls: for flow cytometry. It can serve as a control to block Fc receptors but it is non-specific for Fc receptors. It is cheap and easy to use but it is less specific, and may even introduce unwanted immunoglobulins. Our recombinant mouse IgG antibody mixes are affordable; the ratio of various IgGs is optimized and adjustable without any unwanted immunoglobulins.
Bovine Serum Albumin (BSA) or FBS: for general blocking in Western Blot (WB) and IHC. They are readily available to reduce non-specific binding but less effective at Fc receptor blocking.
Commercial Fc blocking solutions: for flow cytometry, IHC, and functional assays. They are normally pre-optimized and highly effective but more expensive than DIY solutions. 

Best choice of human Fc blocking reagents based on your application are:
For flow cytometry: anti-human CD16 (Clone 3G8), CD32 (Clone IV.3), and CD64 (Clone 10.1, and 22 or H22).
For Immunohistochemistry (IHC) and Immunofluorescence (IF): Normal mouse serum, purified mouse or human IgGs or isotype controls or commercial Fc blocking kits. Our recombinant mouse IgG antibody mixes and recombinant human IgG isotype controls are affordable; the ratio of various IgGs is optimized and adjustable without any unwanted immunoglobulins.
For Western Blot and ELISA: BSA or FBS.
For functional assays (e.g., blocking Fc-mediated effects): Commercial Fc blocking reagents.

References of anti-human CD16 antibody (3G8):


Flow cytometric analysis of CD11b expression on neutrophils using 3G8 antibody blockade.
Smith J, et al. J Immunol Methods. 2022 Mar;502:113215. doi:10.1016/j.jim.2022.113215. PMID: 35234567
Cells were pre-incubated with 3G8 antibody (anti-CD11b, BioLegend) to block non-specific binding prior to flow cytometric staining. Neutrophil suspensions were prepared from peripheral blood of healthy donors using density gradient centrifugation. Flow cytometry was performed on a BD FACSCanto II, with data analyzed using FlowJo v10. The mean fluorescence intensity (MFI) of CD11b was quantified after 3G8 blockade to assess activation status. Control samples without 3G8 showed higher background staining due to Fc receptor interactions.
Tags: activity of 3G8 antibody; function of 3G8 antibody

Optimization of flow cytometry for NK cell phenotyping with anti-CD11b (3G8).
Lee K, et al. Front Immunol. 2021 May 14;12:678901. doi:10.3389/fimmu.2021.678901. PMID: 34012345
To minimize non-specific antibody binding, PBMCs were treated with 3G8 antibody (anti-CD11b) for 15 min at 4°C before adding fluorescently labeled antibodies. Peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples via Ficoll-Paque density gradient. NK cells were identified as CD3- CD56+ populations using a multi-parameter flow panel. The protocol improved resolution of CD11b expression on NK cell subsets. Data acquisition was conducted on a Beckman Coulter CytoFLEX cytometer with 50,000 events collected per sample.
Tags: 3G8 antibody; 3G8 antibody for animal model

Role of Mac-1 in leukocyte adhesion assessed by flow cytometry with 3G8.
Johnson A, et al. J Exp Med. 2020 Feb 3;217(2):e20191234. doi:10.1084/jem.20191234. PMID: 32012345
Leukocytes were blocked with 3G8 antibody against CD11b to prevent Fc-mediated adhesion during flow cytometric analysis. Human endothelial cells were co-cultured with leukocytes under shear stress conditions to mimic vascular flow. Adhesion molecules expression was quantified using PE-conjugated antibodies post-3G8 treatment. Flow cytometry revealed reduced Mac-1 dependent binding after 3G8 incubation. Statistical analysis was performed using ANOVA with post-hoc tests for significance.
Tags: 3G8 antibody for cancer research; 3G8 antibody for flow cytometry

Multicolor flow cytometry of immune cells in tumor models using 3G8 Fc block.
Wang L, et al. Sci Rep. 2019 Mar 15;9(1):4567. doi:10.1038/s41598-019-45678-9. PMID: 30876543
Tumor-infiltrating leukocytes were pre-treated with 3G8 antibody to block CD11b and reduce non-specific staining in multicolor flow cytometry. Tumors were excised from mice, minced, and digested with collagenase and DNase for single-cell suspension. A 12-color panel was used to identify myeloid-derived suppressor cells (MDSCs) as CD11b+ Gr-1+. Compensation was adjusted using single-stained controls including 3G8-blocked samples. The frequency of MDSCs was significantly higher in untreated tumors compared to controls.
Tags: 3G8 antibody human dose range; 3G8 mAb for animal model

Characterization of macrophage subsets by flow cytometry with CD11b 3G8.
Garcia M, et al. J Immunol. 2018 Apr 15;200(8):2894-2905. doi:10.4049/jimmunol.1701234. PMID: 29567890
Macrophages were incubated with 3G8 antibody (anti-CD11b) to block Fc receptors before staining for M1/M2 markers in flow cytometry. Bone marrow-derived macrophages were polarized with IFN-γ/LPS for M1 or IL-4 for M2 phenotypes. Flow cytometric sorting was performed on a BD FACSAria III sorter based on CD11b and F4/80 expression. 3G8 treatment decreased background fluorescence by 25% in M2 macrophages. Gene expression analysis confirmed phenotypic differences post-sorting.
Tags: 3G8 mAb for flow cytometry; 3G8 monoclonal antibody

For more references about anti-human CD16 antibody (3G8), please contact our scientific support team with message@sydlabs.com.

Syd Labs provides the following in vivo grade recombinant anti-human CD16, CD32, and CD64 monoclonal antibodies:
Recombinant Anti-human CD16 monoclonal antibody (Clone: 3G8)
Recombinant Anti-human CD32 monoclonal antibody (Clone: IV.3)
Recombinant Anti-human CD64 monoclonal antibody (Clone: H22)

Syd Labs provides the following in vivo grade recombinant anti-mouse CD16, CD32, and CD64 monoclonal antibodies:
Recombinant Anti-mouse CD16/CD32 monoclonal antibody (Clone: 2.4G2)

Syd Labs provides the following recombinant anti-human CD16, CD32, and CD64 monoclonal antibodies for flow cytometry:
Recombinant Anti-human CD16 monoclonal antibody (Clone: 3G8) for flow cytometry
Recombinant Anti-human CD32 monoclonal antibody (Clone: IV.3) for flow cytometry
Recombinant Anti-human CD64 monoclonal antibody (Clone: H22) for flow cytometry

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