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 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 CompBead to ensure that BD Comp beads 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).
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 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 http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The GD2.F4 monoclonal antibody specifically recognizes CD281 which is also known as Toll like receptor 1 (TLR1). CD281 is a type I transmembrane glycoprotein that belongs to the Toll-like receptor (TLR) family. The extracellular domain of TLR1 is comprised of eight leucine-rich repeats (LRR) whereas the cytoplasmic region contains a Toll/IL-1R (TIR) domain. CD281 forms a heterodimeric receptor with CD282 (TLR2) that recognizes some bacterial lipoproteins and triacylated lipopeptides. CD281 is expressed at low levels by monocytes and lower levels by dendritic cells. As confirmed in development of this antibody, CD281 expression is highly variable amongst human leucocyte donors. CD281 serves as an innate immune receptor that participates in recognizing specific pathogen-associated molecular patterns (PAMPs) expressed by certain pathogens and leads to activation of the NF kappa B signaling pathway by responding effector cells.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.
Note: For optimal performance, users are recommended to titrate this conjugate (BV421 Mouse Anti-Human CD281 (TLR1); Cat. No. 566430) to a concentration of 2 µg/test.
Development References (5)
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Akira S. Toll-like receptors and innate immunity. Adv Immunol. 2001; 78:1-56. (Biology). View Reference
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Dasari P, Nicholson IC, Hodge G, Dandie GW, Zola H. Expression of toll-like receptors on B lymphocytes. Cell Immunol. 2005; 236(1-2):140-145. (Clone-specific: Flow cytometry). View Reference
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Nakao Y, Funami K, Kikkawa S, et al. Surface-expressed TLR6 participates in the recognition of diacylated lipopeptide and peptidoglycan in human cells. J Immunol. 2005; 174(3):1566-1573. (Biology). View Reference
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Visintin A, Mazzoni A, Spitzer JH, Wyllie DH, Dower SK, Segal DM.. Regulation of Toll-like receptors in human monocytes and dendritic cells. J Immunol. 2001; 166(1):249-255. (Clone-specific: Flow cytometry). View Reference
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Wyllie DH, Kiss-Toth E, Visintin A, et al. Evidence for an accessory protein function for Toll-like receptor 1 in anti-bacterial responses. J Immunol. 2000; 165(12):7125-7132. (Immunogen). View Reference
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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.