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 433H monoclonal antibody specifically binds to C-C chemokine receptor type 8 (CC-CKR-8 or CCR8) which is also known as CD198, GPRCY6, CY6, CKRL1, CMKBR8, CMKBRL2, or TER1. CCR8 (CD198) is a seven-transmembrane, G-protein-coupled, glycoprotein receptor that belongs to the beta chemokine receptor family. CCR8 is expressed on monocytes, macrophages, eosinophils, Langerhans cells, thymocytes, and vascular smooth muscle cells. It is preferentially expressed on some T cell subsets including skin resident memory T cells, type 2 T-helper (Th2-like) cells, and T regulatory cells. CCR8 is a receptor for the chemokine C-C motif chemokine 1 (CCL1), also known as SCYA1 or I-309, and plays a role in regulating monocyte and T cell chemotaxis and thymocyte apoptosis. CCR8 may play a role in lung T cell recruitment in asthma and can serve as an alternative coreceptor to CD4 for HIV-1 infection. The 433H antibody reportedly neutralized the chemotactic response of cultured T cells to CCL1.
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 (8)
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D'Ambrosio D, Iellem A, Bonecchi R, et al. Selective up-regulation of chemokine receptors CCR4 and CCR8 upon activation of polarized human type 2 Th cells.. J Immunol. 1998; 161(10):5111-5. (Biology). View Reference
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Fox JM, Najarro P, Smith GL, Struyf S, Proost P, Pease JE. Structure/function relationships of CCR8 agonists and antagonists. Amino-terminal extension of CCL1 by a single amino acid generates a partial agonist.. J Biol Chem. 2006; 281(48):36652-61. (Biology). View Reference
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Haque NS, Fallon JT, Pan JJ, Taubman MB, Harpel PC. Chemokine receptor-8 (CCR8) mediates human vascular smooth muscle cell chemotaxis and metalloproteinase-2 secretion.. Blood. 2004; 103(4):1296-304. (Biology). View Reference
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Lee S, Tiffany HL, King L, Murphy PM, Golding H, Zaitseva MB. CCR8 on human thymocytes functions as a human immunodeficiency virus type 1 coreceptor.. J Virol. 2000; 74(15):6946-52. (Biology). View Reference
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Louahed J, Struyf S, Demoulin JB, et al. CCR8-dependent activation of the RAS/MAPK pathway mediates anti-apoptotic activity of I-309/ CCL1 and vMIP-I.. Eur J Immunol. 2003; 33(2):494-501. (Biology). View Reference
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McCully ML, Ladell K, Hakobyan S, Mansel RE, Price DA, Moser B. Epidermis instructs skin homing receptor expression in human T cells.. Blood. 2012; 120(23):4591-8. (Clone-specific: Flow cytometry). View Reference
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Mutalithas K, Guillen C, Raport C, et al. Expression of CCR8 is increased in asthma.. Clin Exp Allergy. 2010; 40(8):1175-85. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunohistochemistry, Inhibition). View Reference
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Pease JE. Is there a role for CCR8 in the pathogenesis of asthma?. Clin Exp Allergy. 2010; 40(8):1110-2. (Clone-specific). 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.