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RB744 Rat Anti-Mouse CD370 (CLEC9a)

BD OptiBuild™ RB744 Rat Anti-Mouse CD370 (CLEC9a)

Clone 7H11 (also known as No.5; Antibody No.5; Antibody No. 5; Clone 5)

(RUO)
Product Details
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BD OptiBuild™
CD370; Clec9a; Clec-9a; DNGR-1
Mouse (Tested in Development)
Rat Wistar IgG1, κ
Mouse Clec9a-Transfected Rat RBL-2H3 Cells
Flow cytometry (Qualified)
0.2 mg/ml
232414
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.

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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  4. 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.
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. 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.
  7. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  8. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. An isotype control should be used at the same concentration as the antibody of interest.
  11. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
756993 Rev. 1
Antibody Details
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7H11

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.

756993 Rev. 1
Format Details
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RB744
The BD Horizon RealBlue™ 744 (RB744) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).
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RB744
Blue 488 nm
498 nm
746 nm
756993 Rev.1
Citations & References
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View product citations for antibody "756993" on CiteAb

Development References (5)

  1. Cabeza-Cabrerizo M, Cardoso A, Minutti CM, Pereira da Costa M, Reis e Sousa C. Dendritic Cells Revisited.. Annu Rev Immunol. 2021; 39:131-166. (Biology). View Reference
  2. 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
  3. 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
  4. 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
  5. 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
View All (5) View Less
756993 Rev. 1

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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.