BUV563 Mouse Anti-Human CD226
Clone DX11 (RUO)
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- Alternative Name DNAM-1; DNAM1; PTA1; PTA-1; TLiSA1
- Concentration 0.2 mg/ml
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (Tested in Development)
Flow cytometry (Qualified)
- Immunogen Human CTL Clone
- Workshop No. VII 70648
- Entrez Gene ID 10666
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The DX11 monoclonal antibody specifically binds to CD226 which is also known as DNAX accessory molecule-1 (DNAM-1), Platelet and T cell activation antigen 1 (PTA1), or T lineage-specific activation antigen 1 antigen (TLiSA1). CD226 is a 65 kDa type 1 transmembrane glycoprotein consisting of 318 amino acid residues including two Ig-like domains. CD226 is expressed on the majority of T cells, NK cells, monocytes, platelets, and a subset of B cells, but not on erythrocytes. It is also present on a subset of thymocytes coexpressing high density surface CD3. CD226 is not present on normal fibroblast cell lines or tumor cell lines of epithelial or neuronal origins. CD226 is a tyrosine phosphorylated, signal-transducing molecule which participates in primary adhesion during cytotoxic T lymphocyte (CTL)- or NK cell-mediated cytotoxicity. The DX11 antibody inhibits T- and NK cell-mediated cytotoxicity against a variety of tumor cell targets, and blocks cytokine production by alloantigen-specific T cells.
The antibody was conjugated to BD Horizon BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
BUV563 is a tandem fluorochrome that combines BD Horizon BUV395 and an acceptor dye with an EM Max at 563 nm. Due to the excitation of the acceptor dye by other laser lines, there may be spillover into channels detecting PE (eg, 575/26-nm filter) and PE-CF594 (eg, 610/20-nm filter). BUV563 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 BUV563 under optimal conditions that minimize unconjugated dye and antibody.
- This antibody was developed for use in flow cytometry.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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 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.
- CF™ is a trademark of Biotium, Inc.
- BD Horizon Brilliant Ultraviolet 563 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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).