Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) 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).
Product Notices
- 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.
- 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.
- BD Horizon Brilliant Ultraviolet 661 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 UP-D1 monoclonal antibody specifically binds to Immune receptor expressed on myeloid cells 1 (IREM-1). IREM-1 is encoded by CD300LF. It is also known as CD300 antigen-like family member F (CD300f), CMRF35-like molecule 1 (CLM-1/CLM1), NK inhibitory receptor (NKIR), or Immunoglobulin superfamily member 13 (IgSF13). IREM-1/CD300f is expressed on monocytes, dendritic cells, granulocytes, and mast cells. IREM-1 is a single-pass type I transmembrane glycoprotein that belongs to the CMRF family of the Ig gene superfamily. The long intracellular region of IREM-1 contains two classical immunoreceptor tyrosine-based inhibitory motifs (ITIM). Ligand-bound IREM-1 can transduce negative regulatory signals intracellularly by recruiting SHP1 or SHIP phosphatases that in turn dephosphorylate tyrosine residues within activating cell signaling molecules. CD300f may thus play a role in regulating inflammatory responses involving myeloid cells.
The antibody was conjugated to BD Horizon™ BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP. Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, 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.
Development References (6)
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Aguilar H, Alvarez-Errico D, García-Montero AC, Orfao A, Sayós J, López-Botet M. Molecular characterization of a novel immune receptor restricted to the monocytic lineage. J Immunol. 2004; 173(11):6703-6711. (Biology). View Reference
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Alvarez-Errico D, Aguilar H, Kitzig F, Brckalo T, Sayós J, López-Botet M. IREM-1 is a novel inhibitory receptor expressed by myeloid cells. Eur J Immunol. 2004; 34(12):3690-3701. (Immunogen: ELISA, Flow cytometry, Functional assay, Immunoprecipitation, Inhibition). View Reference
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Alvarez-Errico D, Sayós J, López-Botet M. The IREM-1 (CD300f) inhibitory receptor associates with the p85alpha subunit of phosphoinositide 3-kinase. J Immunol. 2007; 178(2):808-816. (Clone-specific: Flow cytometry, Functional assay, Inhibition). View Reference
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Borrego F. The CD300 molecules: an emerging family of regulators of the immune system. Blood. 2013; 121(11):1951-1960. (Biology). View Reference
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Lee SM, Suk K, Lee WH. Synthetic peptides containing ITIM-like sequences of IREM-1 (CD300F) differentially regulate MyD88 and TRIF-mediated TLR signalling through activation of SHP and/or PI3K. Clin Exp Immunol. 2012; 167(3):438-446. (Biology). View Reference
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Sui L, Li N, Liu Q, et al. IgSF13, a novel human inhibitory receptor of the immunoglobulin superfamily, is preferentially expressed in dendritic cells and monocytes. Biochem Biophys Res Commun. 2004; 319(3):920-928. (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.