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PE Rat Anti-Mouse CD370 (Clec9a)

PE Rat Anti-Mouse CD370 (Clec9a)

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

(RUO)
PE Rat Anti-Mouse CD370 (Clec9a)

Multicolor flow cytometric analysis of mouse CD370 (Clec9A) expression on splenic leucocytes. BALB/c mouse splenocytes were stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553729), BD Horizon™ BUV395 Rat Anti-Mouse CD8a (Cat. No. 563786/565968), and APC Hamster Anti-Mouse CD11c (Cat. No. 550261) antibodies and with either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plot) or PE Rat Anti-Mouse CD370 (Clec9A) antibody (Cat. No. 567620; Right Plot) at 0.25 µg/test. A bivariate pseudocolor density plot showing the correlated expression of CD370 (Clec9a) [or Ig Isotype control staining] versus CD11c was generated from CD8+CD4- gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.

Multicolor flow cytometric analysis of mouse CD370 (Clec9A) expression on splenic leucocytes. BALB/c mouse splenocytes were stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553729), BD Horizon™ BUV395 Rat Anti-Mouse CD8a (Cat. No. 563786/565968), and APC Hamster Anti-Mouse CD11c (Cat. No. 550261) antibodies and with either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plot) or PE Rat Anti-Mouse CD370 (Clec9A) antibody (Cat. No. 567620; Right Plot) at 0.25 µg/test. A bivariate pseudocolor density plot showing the correlated expression of CD370 (Clec9a) [or Ig Isotype control staining] versus CD11c was generated from CD8+CD4- gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
CD370; Clec9a; Clec-9a; DNGR-1
Mouse (QC Testing)
Rat Wistar IgG1, κ
Mouse Clec9a-Transfected Rat RBL-2H3 Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
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 and unconjugated antibody and free dye were removed.

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. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. An isotype control should be used at the same concentration as the antibody of interest.
567620 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.

        

567620 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
567620 Rev.1
Citations & References
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Development References (5)

  1. 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
  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
567620 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


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For Research Use Only. Not for use in diagnostic or therapeutic procedures.