BUV496 Rat Anti-Mouse CD370 (Clec9a)
Clone 7H11 (also known as No.5; Antibody No.5; Antibody No. 5; Clone 5)
(RUO)
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).
- BD Horizon Brilliant Ultraviolet 496 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 7H11 monoclonal antibody specifically recognizes mouse CD370 which is also known as DNGR-1 (Dendritic cell natural killer lectin group receptor 1). CD370 is an ~27 kDa single-pass type II transmembrane glycoprotein that is encoded by Clec9A (C-type lectin domain family member 9A) which belongs to the C-type lectin superfamily. CD370 (Clec9a) is selectively expressed as a disulfide-linked homodimer on plasmacytoid dendritic cells and CD8+ myeloid dendritic cells. This receptor is comprised of an extracellular region that contains a C-type lectin domain with a single carbohydrate recognition domain (CRD), followed by a transmembrane segment and a cytoplasmic tail with an immunoreceptor tyrosine-based activation motif (ITAM). CD370 (Clec9a) acts as a damage-associated molecular pattern receptor, ie, an endocytic receptor for damage-associated molecular patterns (DAMPs) that are expressed by necrotic cells, eg, cells dying due to infection or transformation. This receptor binds by its extracellular domain to exposed F-actin-myosin complexes of dead cells. Intracellular signaling mediated through ligand-bound CD370 (Clec9a) leads to phosphorylation of Syk tyrosine kinase. Further downstream signaling can foster MHC Class I-mediated cross-presentation of dead-cell associated antigens by dendritic cells to CD8+ T cells.
The antibody was conjugated to BD Horizon™ BUV496 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 496-nm. BD Horizon BUV496 can be excited by the ultraviolet laser (355 nm) and detected with a 515/30 nm filter with a 450LP. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into the channel detecting BD Horizon V500 or BV510 (eg, 525/40-nm filter). However, the spillover can be corrected through compensation as with any other dye combination.
Development References (5)
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Cabeza-Cabrerizo M, Cardoso A, Minutti CM, Pereira da Costa M, Reis ESC. Dendritic Cells Revisited. Annu Rev Immunol. 2021; 39:1-36. (Biology). View Reference
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Canton J, Blees H, Henry CM, et al. The receptor DNGR-1 signals for phagosomal rupture to promote cross-presentation of dead-cell-associated antigens. Nat Immunol. 2021; 22(2):140-153. (Clone-specific: Flow cytometry). View Reference
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Hanč P, Fujii T, Iborra S, et al. Structure of the Complex of F-Actin and DNGR-1, a C-Type Lectin Receptor Involved in Dendritic Cell Cross-Presentation of Dead Cell-Associated Antigens.. Immunity. 2015; 42(5):839-849. (Clone-specific: Flow cytometry, Functional assay). View Reference
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Hossain MK, Wall KA. Use of Dendritic Cell Receptors as Targets for Enhancing Anti-Cancer Immune Responses.. Cancers (Basel). 2019; 11(3):E418. (Biology). View Reference
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Sancho D, Mourão-Sá D, Joffre OP, et al. Tumor therapy in mice via antigen targeting to a novel, DC-restricted C-type lectin. J Clin Invest. 2008; 118(6):2098-2110. (Immunogen: Flow cytometry, Functional assay, Immunofluorescence). View Reference
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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.
