Catalog No.	PA007277.m1
Product Name	Anti-human CD16 Antibody (Clone: 3G8) | PA007277.m1
Supplier Name	Syd Labs, Inc.
Brand Name	Syd Labs
Clone	3G8.
Isotype	Mouse IgG1 kappa
Source/Host	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	Anti-human CD16 Monoclonal Antibody (Clone: 3G8) 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.
Note	The human CD16 Fc blocking antibody (3G8 Fc blocking antibody) is an anti-human Fc receptor antibody for human FcR blocking reagent. Various isotypes are available for certain applications. Condition of sample preparation and optimal sample dilution should be determined experimentally by the investigator.
Order Offline	Phone: 1-617-401-8149 Fax: 1-617-606-5022 Email: message@sydlabs.com Or leave a message with a formal purchase order (PO) Or credit card.

Description

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

Recombinant mouse IgG1 Monoclonal Antibody.

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 Monoclonal Antibodies:

1、Runx2 stimulates neoangiogenesis through the Runt domain in melanoma.
Cecconi D, et al. Sci Rep. 2019 May 29;9(1):8052. doi: 10.1038/s41598-019-44552-1. PMID: 31142788.
“In order to further explain the observed expression changes in 3G8 cells, the IPA Upstream Regulator analysis was performed to identify a putative cascade of upstream regulators. …To assess the global effects of Runt domain deletion from RUNX2, a comparative shotgun proteomic analysis of A375 and 3G8 melanoma cells was carried out on two biological replicates. …To validate proteomic data, we performed immunoblot analysis to determine the relative amount of up and down-regulated candidates in 3G8 cells compared to A375 cells. …Overall, IPA software revealed that proteins modulated in 3G8 cells show a significant link with many different pathways. …Therefore, we investigated the expression of CD105 and CD31, the markers of tumour neoangiogenesis, in HUVEC co-cultured with NHEM, A375 or 3G8 melanoma cells.”

2、Identification of a novel immunological epitope on Hexon of fowl adenovirus serotype 4.
Liu J, et al. AMB Express. 2021 Nov 20;11(1):153. doi: 10.1186/s13568-021-01309-2. PMID: 34800173.
“Then, mAb 3G8 diluted at 1:1000 was added to the cells and incubated for 30 min at room temperature. …To identify the antigenic epitope of Hexon protein recognized by mAb 3G8, a number of truncated Hexon fragments were amplified using primers listed in Table. …After blocking with 5% skimmed milk in TBST for 2 h at room temperature, the membrane was incubated with mAb 3G8 diluted at 1:1000 in TBST at 37 °C for 1 h. …A hybridoma cell line secreting mAb, designated as 3G8, was generated. …To confirm the amino acid alignment results, we evaluated the reaction of mAb 3G8 with different serotypes of FAdVs.”

3、The impact of N-glycan conformational entropy on the binding affinity of Fc γ receptor IIIa/CD16a.
Kremer PG, et al. Structure. 2025 Dec 19:S0969-2126(25)00475-7. doi: 10.1016/j.str.2025.11.015. PMID: 41421343.
“The antibody 3G8 originated from a mouse immunized with human neutrophils following sorting to identify antibodies that competed with IgG1 for neutrophil binding. …Thus, a structural explanation of the 3G8 Fab interaction with FcγRIII could promote the development of pan-FcγRIII reactive molecules. …We explored the structures of 3G8 and IgG1 Fc binding to the F158 allotype and estimated the energetic impact of glycan conformational entropy on IgG1 Fc binding affinity. …Though it is known that 3G8 binds to cells expressing FcγRIIIa F158 or V158, the affinity of these two complexes was not known. …The S160 residue appears to form a hydrogen bond with the 3G8 residue R58, but the S160A mutation did not reduce affinity, and thus S160A likely contributes negligible stabilizing energy.”

4、Comparison of the different anti-CD16 antibody clones in the activation and expansion of peripheral blood NK cells.
Kim J, et al. Sci Rep. 2023 Jun 11;13(1):9493. doi: 10.1038/s41598-023-36200-6. PMID: 37302991.
“In this study, we evaluated the differences in NK cell response and expansion of four anti-CD16 antibody clones, namely CB16, 3G8, B73.1, and MEM-154, in the same NK cell expansion system. …Numerous monoclonal antibodies have been developed against CD16. In fact, anti-CD16 clones 3G8 and B73.1 have been commonly used to stimulate NK cells in previous studies. …Although we could not demonstrate it in this study, it is possible that the variation in NK cell expansion in response to CB16 clone stimulation among donors is associated with CD16 polymorphisms, similar to what is known for the binding of 3G8 clone to CD16. …All experiments were performed in accordance with relevant guidelines and regulations. …After day 7, the concentration of IL-2 increased from 10 U/mL to 100 U/mL, and 5 ng/mL soluble IL-15 was added to the medium.”

5、Highly Sensitive and Practical Fluorescent Sandwich ELISA for Ciguatoxins.
Tsumuraya T, et al. Anal Chem. 2018 Jun 19;90(12):7318-7324. doi: 10.1021/acs.analchem.8b00519. PMID: 29770692.
“The ELISA protocol, using one microtiter plate coated with two mAbs (10C9 and 3G8), and ALP-linked 8H4, can detect any of the four CTX congeners in a single operation. …CTX1B spiked into fish at the FDA guidance level of 0.01 ppb CTX1B equivalent toxicity in fish from Pacific regions was also proven to be reliably detected by this ELISA. …Ciguatera fish poisoning (CFP) caused by the consumption of fish that have accumulated ciguatoxins (CTXs) affects more than 50000 people annually. …The difficulty in avoiding CFP arises from the lack of sensitive and reliable analytical methods for the detection and quantification of CTXs in contaminated fish, along with the normal appearance, smell, and taste of fish contaminated with the causative toxins. …Thus, an accurate, sensitive, routine, and portable detection method for CTXs is urgently required.”

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

Anti-human CD16 (3G8) from: Recombinant Anti-human CD16 Monoclonal Antibody (Clone: 3G8), Mouse IgG1 Kappa: PA007277.m1 Syd Labs

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