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PE Mouse Anti-Mouse CD289 (TLR9)
PE Mouse Anti-Mouse CD289 (TLR9)

Multicolor flow cytometric analysis of TLR9 (CD289) expression in mouse splenocytes. Mouse splenic leucocytes were stained with Fixable Viability Stain 450 (FVS450; Cat. No. 562247), fixed, and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The cells were then stained with FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 553087/553088/561877) and APC Hamster Anti-Mouse CD11c (Cat. No. 550261/561119) antibodies, and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dotted line histograms) or PE Mouse Anti-Mouse TLR9 antibody (Cat. No. 565640; solid line histograms).  The fluorescence histograms (Bottom Plots) showing total cellular TLR9 (CD289) expression [or Ig Isotype control staining] for the different leucocyte subsets were derived from CD11c versus CD45R/B220 coexpression-gated events with the forward and side light-scatter (SSC-A) characteristics of intact FVS450 live cell-discriminated leucocyte subsets (Top Plots). Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Multicolor flow cytometric analysis of TLR9 (CD289) expression in mouse splenocytes. Mouse splenic leucocytes were stained with Fixable Viability Stain 450 (FVS450; Cat. No. 562247), fixed, and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The cells were then stained with FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 553087/553088/561877) and APC Hamster Anti-Mouse CD11c (Cat. No. 550261/561119) antibodies, and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dotted line histograms) or PE Mouse Anti-Mouse TLR9 antibody (Cat. No. 565640; solid line histograms).  The fluorescence histograms (Bottom Plots) showing total cellular TLR9 (CD289) expression [or Ig Isotype control staining] for the different leucocyte subsets were derived from CD11c versus CD45R/B220 coexpression-gated events with the forward and side light-scatter (SSC-A) characteristics of intact FVS450 live cell-discriminated leucocyte subsets (Top Plots). Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
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BD Pharmingen™
TLR9; Tlr9; Toll-like receptor 9; CD289
Mouse (QC Testing)
Mouse IgG1, κ
Mouse TLR9 Transfected Cell Line
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2739316
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

  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
565640 Rev. 1
Antibody Details
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J15A7

The J15A7 monoclonal antibody specifically binds to the mouse Toll-like receptor 9 (TLR9), which is also known as CD289. It recognizes the TLR9N fragment of mouse TLR9 and does not crossreact with human TLR9. TLR9 is a type I transmembrane protein that belongs to the TLR family whose members play fundamental roles in pathogen recognition and activation of innate immunity. TLR9 recognizes unmethylated CpG sequences in DNA molecules.   It is expressed by numerous cells of the immune system including B lymphocytes, monocytes, natural killer cells, and subsets of dendritic cells (DC). It is primarily expressed intracellularly within the endosomal compartments. TLR9 functions to alert the immune system of viral and bacterial infections by binding to microbial DNA that is rich in CpG motifs. TLR9 signaling leads to the activation of cells that produce proinflammatory cytokines, such as type-I interferon and IL-12. It can also help induce B cell proliferative responses. TLR9 can reportedly be detected on the surfaces of some DC subsets.

565640 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.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
565640 Rev.1
Citations & References
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Development References (2)

  1. Hemmi H, Takeuchi O, Kawai T, et al. A Toll-like receptor recognizes bacterial DNA. Nature. 2000; 408(6813):740-745. (Biology). View Reference
  2. Onji M, Kanno A, Saitoh S, et al. An essential rolw for the N-terminal fragment of Toll-like receptor 9 in DNA sensing. Nat Commun. 2013; 4:1949. (Immunogen: Flow cytometry, Fluorescence microscopy, Immunofluorescence, Immunoprecipitation). View Reference
565640 Rev. 1

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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.