BD Pharmingen™ Alexa Fluor® 647 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 Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Panel) or Alexa Fluor® 647 Rat Anti-Human Clec9A (CD370) antibody (Cat. No. 564267; 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 Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Panel) or Alexa Fluor® 647 Rat Anti-Human Clec9A (CD370) antibody (Cat. No. 564267; 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
BD Pharmingen™
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_2738717
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 647 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557690
PE Mouse Anti-Human HLA-DR RUO
Size
50 Tests
Cat No.
556644
PE Mouse Anti-Human HLA-DR RUO
Size
100 Tests
Cat No.
555812
PE Mouse Anti-Human HLA-DR RUO
Size
25 Tests
Cat No.
560943
564267 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).
564267 Rev. 2
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
564267 Rev.2
Citations & References
View product citations for antibody "564267" on CiteAb
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
564267 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.