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BV421 Mouse Anti-Human TLR4 (CD284)
BV421 Mouse Anti-Human TLR4 (CD284)
Multiparameter flow cytometric analysis of TLR4 (CD284) expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse Anti-Human TLR4 (CD284) antibody (Cat. No. 564401; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of TLR4 (CD284) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multiparameter flow cytometric analysis of TLR4 (CD284) expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse Anti-Human TLR4 (CD284) antibody (Cat. No. 564401; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of TLR4 (CD284) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
TLR4; CD284; Toll-like Receptor 4; ARMD10
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human TLR4 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
7099
AB_2738792
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. 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).
  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. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. An isotype control should be used at the same concentration as the antibody of interest.
564401 Rev. 1
Antibody Details
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TF901

The TF901 monoclonal antibody specifically binds to TLR4 (Toll-like receptor 4) which is also known as CD284. TLR4 is a 110 kDa type I transmembrane glycoprotein that belongs to the Toll-like receptor (TLR) family. TLR4 is expressed on monocytes, macrophages, granulocytes, dendritic cells and endothelial cells. TLR4 binds lipopolysaccharide (LPS) which is a major membrane component of Gram-negative bacteria. It thus acts as an innate immune recognition receptor against many pathogens. In association with MD-2, LPS binds to and signals through TLR4 receptors by MyD88-dependent and MyD88-independent pathways to induce the cellular production of proinflammatory cytokines. TF901 antibody binding is not influenced by the human TLR4 D299G/T399I polymorphism. Preincubation of TLR4-positive cells with the TF901 antibody blocks the subsequent binding of the HTA125 antibody which is also specific for human TLR4.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

564401 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
564401 Rev.1
Citations & References
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Development References (7)

  1. Akashi S, Ogata H, Kirikae F, et al. Regulatory roles for CD14 and phosphatidylinositol in the signaling via toll-like receptor 4-MD-2. Biochem Biophys Res Commun. 2000; 268(1):172-177. (Biology). View Reference
  2. Akira S. Toll-like receptors and innate immunity. Adv Immunol. 2001; 78:1-56. (Biology). View Reference
  3. Beutler B. Tlr4: central component of the sole mammalian LPS sensor. Curr Opin Immunol. 2000; 12(1):20-26. (Biology). View Reference
  4. Rock FL, Hardiman G, Timans JC, Kastelein RA, Bazan JF. A family of human receptors structurally related to Drosophila Toll. Proc Natl Acad Sci U S A. 1998; 95(2):588-593. (Biology). View Reference
  5. Shimazu R, Akashi S, Ogata H, et al. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med. 1999; 189(11):1777-1782. (Biology). View Reference
  6. Yamakawa N, Ohto U, Akashi-Takamura S, et al. Human TLR4 polymorphism D299G/T399I alters TLR4/MD-2 conformation and response to a weak ligand monophosphoryl lipid A. Int Immunol. 2013; 25(1):45-52. (Immunogen: Bioassay, Blocking, Flow cytometry, Immunoprecipitation, Inhibition). View Reference
  7. Zarember KA, Godowski PJ. Tissue expression of human Toll-like receptors and differential regulation of Toll-like receptor mRNAs in leukocytes in response to microbes, their products, and cytokines. J Immunol. 2002; 168(2):554-561. (Biology). View Reference
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564401 Rev. 1

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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.