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BV421 Rat Anti-Human Clec9A (CD370)

BD Horizon™ BV421 Rat Anti-Human Clec9A (CD370)

Clone 3A4/Clec9A (also known as 20/05-3A4 or 3A4)

(RUO)
BV421 Rat Anti-Human Clec9A (CD370)
Multicolor flow cytometric analysis of human Clec9a expression on dendritic cell subsets from human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) were stained with either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon BV421 Rat Anti-Human Clec9A (CD370) antibody (Cat. No. 564266; Right Panel). The cells were also stained with PE Mouse Anti-Human HLA-DR (Cat No. 556644/555812/560943) and BD Horizon™ BV711 Mouse Anti-Human CD141 (Cat. No. 563155) antibodies and FITC Lineage Cocktail 1 (LIN 1) - CD3, CD14, CD16, CD19, CD20, CD56 (Cat. No. 340546) - a lineage cocktail comprised of FITC-conjugated antibodies for the specified CD markers. Two-color flow cytometric dot plots showing the correlated expression CD141 versus Clec9A (CD370) (or Ig Isotype control staining) were derived from Lineage-negative, HLA-DR-positive gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of human Clec9a expression on dendritic cell subsets from human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) were stained with either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon BV421 Rat Anti-Human Clec9A (CD370) antibody (Cat. No. 564266; Right Panel). The cells were also stained with PE Mouse Anti-Human HLA-DR (Cat No. 556644/555812/560943) and BD Horizon™ BV711 Mouse Anti-Human CD141 (Cat. No. 563155) antibodies and FITC Lineage Cocktail 1 (LIN 1) - CD3, CD14, CD16, CD19, CD20, CD56 (Cat. No. 340546) - a lineage cocktail comprised of FITC-conjugated antibodies for the specified CD markers. Two-color flow cytometric dot plots showing the correlated expression CD141 versus Clec9A (CD370) (or Ig Isotype control staining) were derived from Lineage-negative, HLA-DR-positive gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
CD370; C-type lectin domain family 9 member A; CLEC9A; DNGR1
Human (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2a, κ
Human Clec9A Peptide
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2738716
Aqueous buffered solution containing BSA and ≤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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
564266 Rev. 2
Antibody Details
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3A4/Clec9A

The 3A4/Clec9A monoclonal antibody specifically binds to human Clec9A (C-type lectin domain family member 9A). Clec9A is a type II transmembrane glycoprotein with a single extracellular C-type lectin domain. Clec9A is also known as CD370 and DNGR1 (Dendritic cell natural killer lectin group receptor 1). Clec9A is selectively expressed on CD141+ myeloid dendritic cells. Clec9A reportedly functions as an endocytic receptor for necrotic cells. It can mediate the cross-presentation of dead-cell associated antigens by dendritic cells in a Syk-dependent manner. The 3A4 antibody reportedly crossreacts with peripheral blood dendritic cells form Pigtail macaques (Macaca nemestrina).

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

564266 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
564266 Rev.2
Citations & References
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Development References (6)

  1. Caminschi L, Proietto AL, Ahmet F, et al. The dendritic cell subtype restricted C-type lectin Clec9A is a target for vaccine enhancement. Blood. 2008; 112(8):3264-3273. (Immunogen: Flow cytometry). View Reference
  2. Clark HF, Gurney AL, Abaya E, et al. The secreted protein discovery initiative (SPDI), a large-scale effort to identify novel human secreted and transmembrane proteins: a bioinformatics assessment. Genome Res. 2003; 13(10):2265-2270. (Biology). View Reference
  3. Huysamen C, Willment JA, Dennehy KM, Brown GD. CLEC9A is a novel activation C-type lectin-like receptor expressed on BDCA3+ dendritic cells and a subset of monocytes. J Biol Chem. 2008; 283(24):16693-16701. (Biology). View Reference
  4. Jongbloed SL, Kassianos AJ, McDonald KJ, et al. Human CD141+ (BDCA-3)+ dendritic cells (DCs) represent a unique myeloid DC subset that cross-presents necrotic cell antigens. J Exp Med. 2010; 207(6):1247-1260. (Clone-specific: Flow cytometry). 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. (Biology). View Reference
  6. Zhang JG, Czabotar PE, Policheni AN, et al. The dendritic cell receptor Clec9A binds damaged cells via exposed actin filaments. Immunity. 2012; 36(4):646-657. (Biology). View Reference
View All (6) View Less
564266 Rev. 2

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