Description
The monoclonal antibody 29A1.4 specifically binds to mouse CD335, also known as NKp46. NKp46 is a 46 kDa type I transmembrane glycoprotein that is a member of the natural cytotoxicity receptor (NCR) family and immunoglobulin superfamily. NKp46 is encoded by the Ncr1 gene located on chromosome 7. NKp46 functions as a cytotoxicity triggering receptor and is selectively expressed by immature and mature NK cells in all mouse strains tested. NKp46 is detected on a minute fraction of NK-like T cells (less than 2% of NKp46+ express CD3e) but not on CD1d-restricted NKT cells from C57BL/6 mice. When immobilized on tissue culture plates, the 29A1.4 antibody reportedly stimulates NK cells to produce interferon-gamma and to release their cytoplasmic granule contents. Although the ligands for the NKp46 receptor have not been fully characterized, recent evidence indicates that this receptor plays an important role in the NK cell-mediated recognition and killing of some virus-infected cells and tumor cells. The immunogen used to generate the 29A1.4 clone was mouse NKp46-Fc recombinant protein.
The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 737 nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 nm filter. Due to the excitation of the acceptor dye by the red laser line, there may be significant spillover into red laser detectors with filters in the 700-720 nm range.
Format
- Format
BUV737
- Excitation Source
Ultraviolet 355 nm
- Excitation Max
348 nm
- Emission Max
737 nm
BUV737 is a tandem fluorochrome that combines BD Horizon BUV395 and an acceptor dye with an Em Max at 737 nm. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover in to channels detecting Alexa Fluor® 700 like dyes (for example, 712/20-nm filter). BUV737 has been exclusively developed by BD Biosciences for instruments equipped with a 355-nm UV laser.
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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 BUV737 under optimum conditions, and unconjugated antibody and free BD Horizon BUV737 were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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 Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome-conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads. This will ensure that BD CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
