Description
The 3G8 monoclonal antibody specifically binds to the 50-65 kDa transmembrane form of the IgG Fc Receptor (FcãRIII), a human NK cell-associated antigen. CD16 is expressed on NK cells as well as macrophages and granulocytes. Reports indicate that CD16 plays a role in signal transduction and NK cell activation. The 3G8 antibody blocks the binding of soluble immune complexes to granulocytes. The 3G8 antibody is reported (Vossebeld et al., 1997) to increase intracellular calcium levels in human neutrophils by interacting with both FcγRIIa and FcγRIIIb molecules. This antibody has also been reported to induce homotypic neutrophil aggregation.
This clone also cross-reacts with a subset of peripheral blood lymphocytes and monocytes, but not granulocytes, of baboon and both rhesus and cynomologus macaque monkeys. Multi-color analysis reveals that the distribution on lymphocytes is similar to that found in human studies with the majority of CD16-positive lymphocytes being both CD3 and CD20 negative.
The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is
available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.
When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls. Due to spectral differences between cells and beads in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
Format
- Format
V500
- Excitation Source
Violet 405 nm
- Excitation Max
415 nm
- Emission Max
500 nm
BD Horizon™ V500 is a novel organic dye excited by the violet laser. This dye offers improved brightness over Pacific Orange™ and reduced spillover into the FITC channel when compared to AmCyan. BD Horizon V500 cannot be used simultaneously with AmCyan or Pacific Orange™.
Suggested Companion Products
Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with BD Horizon™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- BD Horizon V500 is covered by one or more of the following US patents: 8,431,416.
- Species testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
