BD Pharmingen™ PE Rat Anti-Human Clec9A (CD370)
Clone 3A4/Clec9A (also known as 20/05-3A4 or 3A4)
(RUO)
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 PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Panel) or PE Rat Anti-Human Clec9A (CD370) antibody (Cat. No. 563488; Right Panel). The cells were also stained with FITC Mouse Anti-Human HLA-DR (Cat No. 555811), BD Horizon™ BV711 Mouse Anti-Human CD141 (Cat. No. 563155) and a Lineage cocktail comprised of BD Horizon™ V450-conjugated Mouse Anti-Human CD3 (Cat. No. 560366), Mouse Anti-Human CD14 (Cat. No. 560350), Mouse Anti-Human CD19 (Cat. No. 560354) and Mouse Anti-Human CD56 (Cat. No. 560361) antibodies. Two-color flow cytometric dot plots show the correlated expression patterns of Clec9A (CD370) (or Ig Isotype control staining) versus CD141 for 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 LSRFortessa™ Flow Cytometer System.
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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 PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Panel) or PE Rat Anti-Human Clec9A (CD370) antibody (Cat. No. 563488; Right Panel). The cells were also stained with FITC Mouse Anti-Human HLA-DR (Cat No. 555811), BD Horizon™ BV711 Mouse Anti-Human CD141 (Cat. No. 563155) and a Lineage cocktail comprised of BD Horizon™ V450-conjugated Mouse Anti-Human CD3 (Cat. No. 560366), Mouse Anti-Human CD14 (Cat. No. 560350), Mouse Anti-Human CD19 (Cat. No. 560354) and Mouse Anti-Human CD56 (Cat. No. 560361) antibodies. Two-color flow cytometric dot plots show the correlated expression patterns of Clec9A (CD370) (or Ig Isotype control staining) versus CD141 for 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 LSRFortessa™ Flow Cytometer System.
Product Details
BD Pharmingen™
CD370; C-type lectin domain family 9 member A; CLEC9A; DNGR1
Human (QC Testing), Monkey (Reported)
Rat WI, also known as Wistar (outbred) IgG2a, κ
Human Clec9A Peptide
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2738237
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
PE Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
553930
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FITC Mouse Anti-Human HLA-DR RUO
Size
100 Tests
Cat No.
555811
BV711 Mouse Anti-Human CD141 RUO
Size
50 Tests
Cat No.
563155
V450 Mouse Anti-Human CD3 RUO
Size
30 Tests
Cat No.
560366
V450 Mouse Anti-Human CD14 RUO
Size
30 Tests
Cat No.
560350
563488 Rev. 2
Antibody Details
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).
563488 Rev. 2
Format Details
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.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
563488 Rev.2
Citations & References
Development References (6)
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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
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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
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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
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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
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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. (Biology). View Reference
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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
563488 Rev. 2
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.
