Multiparameter flow cytometric analysis of TLR5 (CD285) expression on human peripheral blood leucocytes. Whole blood was stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 555397/557153/561712) and either Alexa Fluor® 647 mouse IgG1, κ Isotype Control [Cat. No. 557714; Left Panels] or Alexa Fluor® 647 Mouse Anti-Human TLR5 (CD285) antibody [Cat. No. 564344; Right Panels]. The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots showing the correlated expression of CD14 [Upper Panels] versus TLR5 (CD285) [or Ig Isotype control staining] were derived from gated events with the light-scatter characteristics of viable mononuclear leucocytes. Two-parameter flow cytometric contour plots showing the correlated Side Light Scatter (SSC) characteristics [Lower Panels] versus TLR5 (CD285) expression were derived from gated events with the light-scatter characteristics of total viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Multiparameter flow cytometric analysis of TLR5 (CD285) expression on human peripheral blood leucocytes. Whole blood was stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 555397/557153/561712) and either Alexa Fluor® 647 mouse IgG1, κ Isotype Control [Cat. No. 557714; Left Panels] or Alexa Fluor® 647 Mouse Anti-Human TLR5 (CD285) antibody [Cat. No. 564344; Right Panels]. The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots showing the correlated expression of CD14 [Upper Panels] versus TLR5 (CD285) [or Ig Isotype control staining] were derived from gated events with the light-scatter characteristics of viable mononuclear leucocytes. Two-parameter flow cytometric contour plots showing the correlated Side Light Scatter (SSC) characteristics [Lower Panels] versus TLR5 (CD285) expression were derived from gated events with the light-scatter characteristics of total viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
Toll-like receptor 5; MELIOS; SLEB1; TIL3
Human (QC Testing)
Mouse IgG1, κ
Human TLR5 Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
AB_2738757
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
FITC Mouse Anti-Human CD14 RUO
Size
100 Tests
Cat No.
555397
564344 Rev. 2
Antibody Details
624915
The 624915 monoclonal antibody specifically binds to TLR5 (Toll-like Receptor 5) which is also known as CD285, MELIOS, SLEB1, and TIL3. TLR5 is a type I transmembrane glycoprotein of the Toll-like Receptor family. It is expressed on mucosal epithelial cells and leucocytes, including monocytes and some immature dendritic cells and CD4-positive T cells. CD285 binds to bacterial flagellin. Flagellin binding induces TLR5 dimerization and intracellular signaling through MyD88 resulting in the cellular production of proinflammatory cytokines and immune system activation.
564344 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 520-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
564344 Rev.2
Citations & References
Development References (3)
-
Aderem A, Ulevitch RJ. Toll-like receptors in the induction of the innate immune response. Nature. 2000; 406(6797):782-787. (Biology). View Reference
-
Bates JT, Graff AH, Phipps JP, Grayson JM, Mizel SB. Enhanced antigen processing of flagellin fusion proteins promotes the antigen-specific CD8+ T cell response independently of TLR5 and MyD88. J Immunol. 2011; 186(11):6255-6262. (Biology). View Reference
-
Smith KD, Andersen-Nissen E, Hayashi F, et al. Toll-like receptor 5 recognizes a conserved site on flagellin required for protofilament formation and bacterial motility. Nat Immunol. 2003; 4(12):1247-1253. (Biology). View Reference
564344 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.
