Preparation And Storage
Recommended Assay Procedures
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 CompBead to 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).
Product Notices
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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 http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- BD Horizon Brilliant Ultraviolet 615 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.
Companion Products
The C65-485 monoclonal antibody specifically recognizes the type II membrane protein known as 4-1BB Ligand (4-1BB-L) or CD137 Ligand (CD137L). This molecule belongs to the TNF superfamily and is encoded by TNFSF9 (Tumor necrosis factor superfamily member 9). CD137 Ligand has been reported to have costimulatory function on activated T cells. Reports suggest that stimulation of activated T cells via CD137 Ligand does not require the presence of other costimulatory molecules, including CD28. Some studies have reported human CD137 Ligand to be expressed preferentially on primary B cells and B-cell lines. The C65-485 antibody is able to react with recombinant human 4-1BB Ligand (rh4-1BB-L/CD137L) bound to CD137 (4-1BB receptor) expressed on phytohemagglutinin (PHA)-stimulated PBMC.
The antibody was conjugated to BD Horizon BUV615 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 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP. Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).
Development References (7)
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Chu NR, DeBenedette MA, Stiernholm BJ, Barber BH, Watts TH. Role of IL-12 and 4-1BB ligand in cytokine production by CD28+ and CD28- T cells. J Immunol. 1997; 158(7):3081-3089. (Biology). View Reference
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Cole SL, Benam KH, McMichael AJ, Ho LP. Involvement of the 4-1BB/4-1BBL pathway in control of monocyte numbers by invariant NKT cells.. J Immunol. 2014; 192(8):3898-907. (Clone-specific: Functional assay). View Reference
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DeBenedette MA, Shahinian A, Mak TW, Watts TH. Costimulation of CD28- T lymphocytes by 4-1BB ligand. J Immunol. 1997; 158(2):551-559. (Biology). View Reference
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Ju S, Ju S, Ge Y, et al. A novel approach to induce human DCs from monocytes by triggering 4-1BBL reverse signaling.. Int Immunol. 2009; 21(10):1135-44. (Clone-specific: Flow cytometry). View Reference
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Melero I, Bach N, Hellström KE, Aruffo A, Mittler RS, Chen L. Amplification of tumor immunity by gene transfer of the co-stimulatory 4-1BB ligand: synergy with the CD28 co-stimulatory pathway. Eur J Immunol. 1998; 28(3):1116-1121. (Biology). View Reference
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Pichler K, Kattan T, Gentzsch J, et al. Strong induction of 4-1BB, a growth and survival promoting costimulatory receptor, in HTLV-1-infected cultured and patients' T cells by the viral Tax oncoprotein.. Blood. 2008; 111(9):4741-51. (Clone-specific: Flow cytometry). View Reference
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Zhou Z, Kim S, Hurtado J, et al. Characterization of human homologue of 4-1BB and its ligand. Immunol Lett. 1995; 45(1-2):67-73. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
