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PE Mouse Anti-Mouse CD283 (TLR3)
PE Mouse Anti-Mouse CD283 (TLR3)

Multicolor flow cytometric analysis of CD283 (TLR3) expression by mouse splenocytes. C57BL/6 mouse splenic leucocytes were surface stained with FITC Hamster Anti-Mouse CD3e (Cat. No. 553062/553061/561827), Alexa Fluor® 700 Mouse Anti-Mouse NK1.1 (Cat. No. 560515), BD Horizon™ BV510 Rat Anti-Mouse Ly-6G and Ly-6C (Cat. No. 563040), and BD Horizon™ BV421 Hamster Anti-Mouse CD11c (Cat. No. 562782) antibodies. The cells were washed, fixed, and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The cells were then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Mouse CD283 (TLR3) antibody (Cat. No. 565984; Right Plot). Two-color contour plots showing the correlated expression CD11c versus CD283 (TLR3) [or Ig Isotype control staining] were derived from Ly-6G and Ly-6C- NK1.1- CD3e- gated events with the forward and side light-scatter characteristics of intact splenic leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Multicolor flow cytometric analysis of CD283 (TLR3) expression by mouse splenocytes. C57BL/6 mouse splenic leucocytes were surface stained with FITC Hamster Anti-Mouse CD3e (Cat. No. 553062/553061/561827), Alexa Fluor® 700 Mouse Anti-Mouse NK1.1 (Cat. No. 560515), BD Horizon™ BV510 Rat Anti-Mouse Ly-6G and Ly-6C (Cat. No. 563040), and BD Horizon™ BV421 Hamster Anti-Mouse CD11c (Cat. No. 562782) antibodies. The cells were washed, fixed, and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The cells were then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Mouse CD283 (TLR3) antibody (Cat. No. 565984; Right Plot). Two-color contour plots showing the correlated expression CD11c versus CD283 (TLR3) [or Ig Isotype control staining] were derived from Ly-6G and Ly-6C- NK1.1- CD3e- gated events with the forward and side light-scatter characteristics of intact splenic leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
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BD Pharmingen™
Tlr3; TLR3; Toll-like receptor 3
Mouse (QC Testing)
Mouse BALB/c IgG1, κ
Mouse TLR3 Transfected Cell Line
Intracellular staining (flow cytometry) (Routinely Tested), Flow cytometry (Reported)
0.2 mg/ml
AB_2739436
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.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

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. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
565984 Rev. 1
Antibody Details
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PaT3

The PaT3 monoclonal antibody specifically binds to the Toll-like receptor 3 (TLR3), which is also known as CD283. CD283 belongs to the mammalian toll-like family of pattern recognition receptors (TLR) which participate in innate immune responses to microbial pathogens. It is a type I transmembrane glycoprotein that has an extracellular domain with leucine-rich repeats and a cytoplasmic domain with homology to the cytoplasmic region of IL-1RI/CD121a. CD283 is differentially expressed on splenic CD8+ dendritic cells, macrophages, and marginal zone B cells. It is expressed on the cell surface as well as intracellular compartments including endolysosomes. As in the case of viral infection, CD283 senses double-stranded RNA (dsRNA) in endolysosomes which subsequently leads to the production of antimicrobial peptides and proinflammatory cytokines by responding cells. CD283 can also be triggered by the synthetic analog, polyinosinic:polycytidylic acid (Poly I:C). The PaT3 monoclonal antibody can reportedly augment TLR3-mediated cellular responses to dsRNA. The PaT3 antibody does not crossreact with human CD283 (TLR3).

        

565984 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
565984 Rev.1
Citations & References
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Development References (3)

  1. Alexopoulou L, Holt AC, Medzhitov R, Flavell RA. Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3.. Nature. 2001; 413(6857):732-8. (Biology). View Reference
  2. Jelinek I, Leonard JN, Price GE, et al. TLR3-specific double-stranded RNA oligonucleotide adjuvants induce dendritic cell cross-presentation, CTL responses, and antiviral protection.. J Immunol. 2011; 186(4):2422-9. (Biology). View Reference
  3. Murakami Y, Fukui R, Motoi Y, et al. Roles of the cleaved N-terminal TLR3 fragment and cell surface TLR3 in double-stranded RNA sensing.. J Immunol. 2014; 193(10):5208-17. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunoprecipitation). View Reference
565984 Rev. 1

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