Anti-Human CD16 Monoclonal Antibodies

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 of Anti-human CD16 Monoclonal Antibody (Clone: 3G8)

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 Monoclonal Antibodies:

Human blood neutrophils generate ROS through FcγR-signaling to mediate protection against febrile P. falciparum malaria.
Ofori, E. A., et al. Commun Biol. 2023 Jul;6(1):743. doi: 10.1038/s42003-023-05118-0. PMID: 37463969
Before incubating with merozoites, cells were treated with antibodies against FcγRI (CD64), FcγII (CD32), and FcγIII (CD16), as described previously in ref. 10. Cells were treated for 30 min. at 37 °C with 10 μg/ml of anti-human CD64; anti-human CD32; anti-human CD16, or a combination of these to block FcγR-I, II, and III, respectively. ...Blood phagocytes, such as neutrophils and monocytes, generate reactive oxygen species (ROS) as a part of host defense response against infections. ...Plasmodium falciparum merozoites opsonized with naturally occurring IgG triggered both intracellular and extracellular ROS generation in blood phagocytes, with neutrophils being the main contributors. ...Using specific inhibitors, we show that both FcγRIIIB and FcγRIIA acted synergistically to induce ROS production in neutrophils, and that NADPH oxidase 2 and the PI3K intracellular signal transduction pathway were involved in this process. ...Here, we used peripheral blood leukocytes (PBLs) to study ROS production intracellularly and extracellularly using freshly purified P. falciparum merozoites and naturally occurring antibodies from samples from well-established longitudinal cohort studies (LCS) performed in Ghana and India.
Tags: activity of 3G8 antibody; function of 3G8 antibody

Neutralizing antibodies from the rare convalescent donors elicited antibody-dependent enhancement of SARS-CoV-2 variants infection.
Mu, S., et al. Front Med (Lausanne). 2022 Oct;9. doi: 10.3389/fmed.2022.952697. PMID: 36341247
As SARS-CoV-2 strains continue to evolve, new variants have been increasing in the current global pandemic. ...Here, we constructed a series of SARS-CoV-2 variants, including engineered double-mutant variants containing an RBD mutation and D614G in the spike protein and natural epidemic variants, and tested whether plasma samples and TOP10 neutralizing antibodies in our laboratory from convalescent patients mediated the ADE effects of these SARS-CoV-2 variants. ...Currently, neutralizing antibody and vaccine strategies have been developed by targeting the SARS-CoV-2 strain identified during the early phase of the pandemic. ...Only one antibody, 55A8, from the same batch of convalescent patients mediated the ADE effects of multiple SARS-CoV-2 variants in vitro, including six double-mutant variants and four epidemic variants, suggesting that ADE activities may be closely related to the antibody itself and the SARS-CoV-2 variants' S proteins. ...These data suggested that potential neutralizing antibodies may need to undergo ADE screening tests for SARS-CoV-2 variants, which should aid in the future design of effective antibody-based therapies.
Tags: 3G8 antibody human dose range; 3G8 mAb for animal model

For more references about Anti-Human CD16 Monoclonal Antibodies 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