iF560 Anti-Human CD16 Antibody

Product Details


Clone3G8
ApplicationFlow Cytometry
ReactivityHuman
FormatiF560
Target NameCD16, FcγRIII
IsotypeMouse IgG1
Antibody TypeMonoclonal
Regulatory StatusRUO
FormulationPhosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 0.2% (w/v) BSA
Protein ConcentrationSupplied at a lot-specific concentration.
Storage & HandlingThe antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Recommended UsageFor flow cytometric staining, it is recommended to use 5 µL of this reagent per 0.5-1.0 million cells in a 100 µL volume. Optimal reagent performance should be determined by titration for each specific application. iF560 has an excitation max at 560 nm and an emission max at 571 nm.
Excitation LaserGreen/Yellow laser (532/561nm)
See All FormatsClone 3G8

Background Information


CD16, also known as Fc gamma receptor III (FcγRIII), is a low-affinity receptor for the Fc region of immunoglobulin G (IgG) and plays a pivotal role in antibody-mediated immune responses. It is expressed primarily on natural killer (NK) cells, neutrophils, monocytes, and macrophages, with expression patterns and function varying by cell type. In humans, CD16 exists in two closely related forms encoded by distinct genes: CD16a (FcγRIIIA) and CD16b (FcγRIIIB).

Structurally, CD16 is a type I transmembrane glycoprotein composed of two extracellular immunoglobulin-like domains responsible for IgG binding. CD16a is a transmembrane receptor expressed on NK cells and some myeloid cells, where it associates with signaling adaptor proteins containing immunoreceptor tyrosine-based activation motifs (ITAMs), such as FcεRIγ and CD3ζ. In contrast, CD16b is attached to the cell surface via a glycosylphosphatidylinositol (GPI) anchor and is expressed almost exclusively on neutrophils, lacking direct intracellular signaling capacity.

Functionally, CD16 mediates antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis. Upon binding to IgG-opsonized target cells, CD16a on NK cells triggers activation signals that lead to the release of cytotoxic granules containing perforin and granzymes, resulting in target cell death. On myeloid cells, CD16 engagement promotes phagocytosis, oxidative burst, and cytokine release, contributing to pathogen clearance and inflammation. CD16 preferentially binds IgG1 and IgG3 subclasses, which are commonly elicited during effective immune responses.

CD16 plays important roles in both protective immunity and disease. Genetic polymorphisms in FCGR3A influence IgG binding affinity and have been associated with susceptibility to infections, autoimmune diseases, and cancer outcomes. Excessive or dysregulated CD16 activation contributes to inflammatory and autoimmune conditions such as rheumatoid arthritis and systemic lupus erythematosus, where immune complexes drive tissue damage. In cancer, CD16 expression and function on NK cells are critical determinants of immune surveillance and therapeutic efficacy.

In therapeutics, CD16 is central to the mechanism of action of many antibody-based drugs. Therapeutic monoclonal antibodies used in oncology, such as those targeting tumor antigens, rely on CD16-mediated ADCC for clinical activity. Engineering antibodies with enhanced Fc affinity for CD16 or developing CD16-engaging bispecific antibodies are active areas of drug development. Additionally, adoptive NK cell therapies often aim to optimize CD16 expression and signaling, highlighting CD16 as a key bridge between antibody recognition and cellular immune effector function.

Isotype Control


iF560 Mouse IgG1 Isotype Control Antibody

Data Sheets


iF560 Anti-Human CD16 Antibody TDS

Related Protocols


Flow Cytometry Protocol

Frequently Asked Questions


How do I determine the optimal antibody concentration/titer for my experiment?
We recommend titrating each new antibody lot on your own cell type and instrument, since optimal concentration depends on cell density, target expression level, and cytometer sensitivity. Suggested starting dilutions or test sizes (e.g., µg or µl per 10^6 cells) are provided on the product page as a starting reference point, not a fixed requirement.

Can these antibodies be used for intracellular or intranuclear staining, or only surface staining?
Intended use (surface, intracellular, or both) is specified per product, indicated under Application. The appropriate Related Protocol is linked for each product. If you need to detect both surface and intracellular markers in the same panel, confirm each antibody's compatibility with your fixation/permeabilization protocol before combining them.

What controls should I use alongside these antibodies?
Matched isotype controls (same host species, isotype, and conjugate) are available for most clones to help distinguish specific binding from background/non-specific staining and are linked on each product page. Unstained, single-color compensation, and fluorescence-minus-one (FMO) controls are also recommended, especially for multicolor panel design.

Can I combine InnoCyto Flow Cytometry antibodies into a multicolor panel?
Yes — antibodies across our catalog are designed to be panel-compatible, and conjugate options are offered specifically to support multiplexing. When building a panel, pair bright fluorophores with low-expression targets, avoid excessive spectral overlap, and confirm compensation is set up correctly for your specific fluorochrome combination. Use our Panel Builder to simplify the process.

Have a product or application question? Consult our FAQs or contact us.