Anti-Human CD16 [3G8] – Purified in vivo GOLD™ Functional Grade
Anti-Human CD16 [3G8] – Purified in vivo GOLD™ Functional Grade
Product No.: C2858
Clone 3G8 Target CD16 Formats AvailableView All Product Type Monoclonal Antibody Alternate Names FcγRIII, Fc Gamma RIII Isotype Mouse IgG1 κ Applications B , CyTOF® , FC , IHC FF , in vivo , IP |
Antibody DetailsProduct DetailsReactive Species Baboon ⋅ Chimpanzee ⋅ Cynomolgus Monkey ⋅ Marmoset ⋅ Pigtailed Macaque ⋅ Rhesus Monkey ⋅ Squirrel Monkey ⋅ Human Host Species Mouse Recommended Isotype Controls Recommended Isotype Controls Recommended Dilution Buffer Immunogen Human PMN cells Product Concentration ≥ 5.0 mg/ml Endotoxin Level < 1.0 EU/mg as determined by the LAL method Purity ≥95% monomer by analytical SEC ⋅ >95% by SDS Page Formulation This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration. Product Preparation Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates. Storage and Handling Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles. Country of Origin USA Shipping Next Day 2-8°C RRIDAB_2829613 Applications and Recommended Usage? Quality Tested by Leinco FC The suggested concentration for this 3G8 antibody for staining cells in flow cytometry is ≤ 2.0 μg per 106 cells in a volume of 100 μl or 100μl of whole blood. Titration of the reagent is recommended for optimal performance for each application. Additional Applications Reported In Literature ? CyTOF® IHC (Frozen) IP B Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity Clone 3G8 recognizes an epitope on human CD16. This clone also cross-reacts with non-human primate CD16.
Background CD16 antibody, 3G8, recognizes human CD16, also known as low-affinity IgG receptor III (FcγRIII). There are two distinct forms of CD16, CD16a (FcγRIIIa) and CD16b (FcγRIIIb). CD16a is a 50-65 kDa heterooligomeric polypeptide-anchored transmembrane protein expressed by NK cells, macrophages, and subsets of monocytes1. CD16b is a 48 kDa monomeric glycosylphosphatidylinositol (GPI)-anchored protein expressed on neutrophils1. Upon binding to the Fc portion of IgG or IgG-antigen complex, both CD16 isoforms induce signaling cascades resulting in multiple functions, including phagocytosis, cytokine release, proliferation, degranulation, and antibody dependent cell-mediated cytotoxicity (ADCC)2. Antigen Distribution CD16 is expressed on NK cells, activated monocytes, macrophages, neutrophils, and placental trophoblasts.
Ligand/Receptor Aggregated IgG, IgG-antigen complex Function Low affinity IgG Fc receptor, phagocytosis, ADCC NCBI Gene Bank ID UniProt.org Research Area Cell Biology . Immunology Leinco Antibody AdvisorPowered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments. Common in vivo applications of clone 3G8 in mice include blocking human FcγRIII (CD16)-mediated antibody-dependent cellular cytotoxicity (ADCC), inhibiting neutrophil phagocytosis, and stimulating NK cell proliferation in mouse models engineered to express human CD16 or with engrafted human immune cells. Key applications and contexts:
Important context:
Summary of main functions in vivo (humanized mouse models):
If clarification is needed, please specify if the focus is on wild-type, transgenic, or engrafted mouse models, as 3G8's application is restricted to contexts where human CD16 is expressed. In the literature, several antibodies and proteins are commonly used alongside the 3G8 antibody, particularly in studies related to immune responses and cell activation. Here are a few examples:
These antibodies and proteins are used in various experimental settings to explore immune mechanisms, cell signaling, and therapeutic applications. Clone 3G8 is a widely used monoclonal antibody against CD16 that has generated substantial scientific insights across multiple functional domains. This antibody has proven instrumental in understanding NK cell biology, neutrophil function, and receptor interactions. Functional Activities and MechanismsClone 3G8 exhibits multiple functional activities that have been extensively documented. The antibody inhibits cytotoxic ability, activates cell signaling, and achieves NK cell depletion in vivo. A particularly important finding is that 3G8 blocks neutrophil phagocytosis while simultaneously stimulating NK cell proliferation. The antibody interacts with both FcγRIIa and FcγRIIIb receptors, causing neutrophil activation and aggregation. This interaction has practical implications for experimental design, as staining in whole blood may cause a reduction in the number of granulocytes or alter their scatter profile. Epitope Recognition and Binding CharacteristicsClone 3G8 recognizes an epitope located on the putative FG loop of the membrane-proximal Ig-like domain of CD16, which represents the major binding site for IgG. Cross-blocking studies have demonstrated that 3G8 recognizes the same epitope as CB16, though 3G8 and B73.1 antibody clones bind distinct epitopes. The antibody shows sensitivity to IgG competition, with binding to CD16a on NK cells being significantly affected by the presence of human IgG. This characteristic distinguishes 3G8 from other clones and reflects its binding site overlap with the Fc region of immunoglobulins. NK Cell Response and ExpansionComparative studies evaluating multiple anti-CD16 antibody clones have revealed important differences in their ability to activate and expand NK cells. When measuring CD107a degranulation and IFN-γ and TNF-α cytokine production, 3G8 demonstrated intermediate effectiveness among four tested monoclonal antibodies, ranking second after CB16 but ahead of B73.1 and MEM-154. The NK cell expansion rate also differed depending on which anti-CD16 antibody was coated on microbeads, with these variations likely attributable to distinct epitope recognition patterns. Cross-Reactivity and ApplicationsClone 3G8 demonstrates remarkable cross-reactivity across multiple non-human primate species, including capuchin monkey, chimpanzee, common marmoset, cynomologous monkey, hamadyras baboon, olive baboon, pigtailed macaque, rhesus, and squirrel monkey. This broad reactivity has made it valuable for comparative immunology studies. The antibody has been successfully employed in flow cytometric analysis, functional assays, immunohistochemical staining of acetone-fixed frozen tissue sections, immunoprecipitation, and blocking of immunoglobulin binding to FcγRIII. Affinity Profiling ApplicationsRecent innovations have utilized 3G8 in combination with other antibody clones for "affinity profiling" approaches. By exploiting differences in binding between 3G8 and B73.1 antibodies, researchers can distinguish NK cells based upon IgG-binding affinity to CD16a using flow cytometry bivariate plots. This method provides the ability to differentiate cell populations with varying affinities for immunoglobulin binding. Historical SignificanceClone 3G8 played a pivotal role in understanding NK cell receptor interactions, revealing how low-affinity Fc receptors mediate critical immune responses. Its development contributed foundational knowledge to the field of natural killer cell biology and antibody-dependent cellular cytotoxicity mechanisms. The dosing regimens of clone 3G8, an anti-human CD16 monoclonal antibody, in mouse models can vary significantly based on the specific application and target cell population being studied. However, detailed information on how these regimens differ across various mouse models is not explicitly provided in the available search results. Here are some general considerations for using clone 3G8 in mouse models:
For precise dosing regimens in specific mouse models, it is essential to consult detailed experimental protocols or publications related to those models. If you're planning experiments, it's advisable to titrate the antibody to find the optimal dose for your particular study conditions. This approach ensures that you achieve the desired effects while minimizing potential side effects or off-target results. References & Citations1. Ravetch JV & Perussia B. (1989) J Exp Med. 170(2):481-497 2. Nimmerjahn F & Ravetch JV (2008) Nat Rev Immunol. 8(1):34-47 Technical ProtocolsCertificate of Analysis |
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