Invitrogen

CD284 (TLR4) Monoclonal Antibody (HTA125), PerCP-eFluor™ 710, eBioscience™

23 Published Figures
17 References
View all (67) TLR4 (CD284) antibodies
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Datasheet
Protocols
Questions & Answers
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Product Details

46-9917-42

Applications
Tested Dilution
Publications

Western Blot (WB)

-
View 1 publication 1 publication

Flow Cytometry (Flow)

5 µL (0.5µg)/test
View 15 publications 15 publications

Neutralization (Neu)

-
View 1 publication 1 publication
Product Specifications

Species Reactivity

Human

Published species

Not Applicable

Host/Isotype

Mouse / IgG2a, kappa

Recommended Isotype Control

Mouse IgG2a kappa Isotype Control (eBM2a), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

HTA125

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

Excitation/Emission Max

482/708 nm View spectra spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2848349

Product Specific Information

Description: The HTA125 monoclonal antibody reacts with human Toll-like receptor 4 (TLR4). So far, at least ten members of the Toll family have been identified in humans. This family of type I transmembrane proteins is characterized by an extracellular domain with leucine-rich repeats and a cytoplasmic domain with homology to the type I IL-1 receptor. Two of these receptors, TLR2 and TLR4, are pattern recognition receptors and signaling molecules in response to bacterial lipoproteins and have been implicated in innate immunity and inflammation. TLR4 physically associates with another molecule called MD-2, and together with CD14, this complex is responsible for LPS recognition and signaling. TLR4 is expressed by peripheral blood monocytes. HTA125 has been reported to immunoprecipitate human TLR4 (~100 kDa) from transfected cells. Most TLR cell surface expression, especially TLR1 and TLR4, occurs at low levels on monocytes and at even lower levels on other cell types including granulocytes and immature dendritic cells (iDC). Furthermore, a relatively high degree of variability in TLR surface expression has been reported among normal donors.

It is highly recommended that for optimal staining of TLR4, whole blood be stained using the lysed whole blood protocol (found in Best Protocols) rather than Ficoll-gradient prepared normal human peripheral blood cells. The use of a density gradient appears to reduce the staining intensity significantly.

Applications Reported: This HTA125 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This HTA125 antibody has been pre-diluted and tested by flow cytometric analysis of cells transfected with human TLR4. This may be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Target Information

TLR4 is member of the Toll like receptor (TLR) family which plays a fundamental role in pathogen recognition and activation of innate immunity. TLRs are highly conserved from Drosophila to humans and share structural and functional similarities. This receptor is most abundantly expressed in placenta, and in myelomonocytic subpopulation of the leukocytes. Mammalian cells respond to LPS by activating TLR4. TLR4 belongs to the multi-protein complex of lipopolysaccharide (LPS) receptor, containing CD14, LY96 and TLR4, and is involved in signal transduction events induced by lipopolysaccharide (LPS) found in most gram-negative bacteria. TLR4 aids in the recognition of pathogen-associated molecular patterns (PAMPs) that are expressed on infectious agents, and mediate the production of cytokines necessary for the development of effective immunity. The various TLRs exhibit different patterns of expression. Mutations in the TLR4 gene have been associated with differences in LPS responsiveness. Also, several transcript variants of the TLR4 gene have been found, but the protein coding potential of most of them is uncertain. TLR4 is expressed by peripheral blood monocytes and a small population of B-cells and is also expressed in human placenta. Studies with TLR4-deficient mice indicate that the main ligand for TLR is lipopolysaccharide. Consequently, these mice also showed increased susceptibility to Gram-negative sepsis.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: CD284; homolog of Drosophila toll; hToll; Toll-like receptor 4

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Gene Aliases: ARMD10; CD284; TLR-4; TLR4; TOLL

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UniProt ID: (Human) O00206

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Entrez Gene ID: (Human) 7099

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Function(s)
lipopolysaccharide binding lipopolysaccharide receptor activity receptor activity transmembrane signaling receptor activity protein binding
Process(es)
activation of MAPK activity toll-like receptor signaling pathway B cell proliferation involved in immune response nitric oxide production involved in inflammatory response regulation of dendritic cell cytokine production MyD88-dependent toll-like receptor signaling pathway MyD88-independent toll-like receptor signaling pathway inflammatory response immune response I-kappaB kinase/NF-kappaB signaling I-kappaB phosphorylation positive regulation of platelet activation positive regulation of gene expression astrocyte development detection of fungus positive regulation of B cell proliferation lipopolysaccharide-mediated signaling pathway response to lipopolysaccharide detection of lipopolysaccharide interferon-gamma production negative regulation of interferon-gamma production negative regulation of interleukin-17 production negative regulation of interleukin-23 production negative regulation of interleukin-6 production negative regulation of tumor necrosis factor production positive regulation of chemokine production positive regulation of interferon-alpha production positive regulation of interferon-beta production positive regulation of interferon-gamma production positive regulation of interleukin-1 production positive regulation of interleukin-10 production positive regulation of interleukin-12 production positive regulation of interleukin-6 production positive regulation of interleukin-8 production positive regulation of tumor necrosis factor production negative regulation of MyD88-independent toll-like receptor signaling pathway toll-like receptor 4 signaling pathway TRIF-dependent toll-like receptor signaling pathway T-helper 1 type immune response macrophage activation positive regulation of NF-kappaB import into nucleus positive regulation of tumor necrosis factor biosynthetic process defense response to bacterium positive regulation of interleukin-12 biosynthetic process innate immune response positive regulation of MHC class II biosynthetic process positive regulation of interferon-beta biosynthetic process positive regulation of interleukin-8 biosynthetic process positive regulation of nitric oxide biosynthetic process negative regulation of osteoclast differentiation positive regulation of transcription from RNA polymerase II promoter positive regulation of JNK cascade interleukin-1 beta secretion regulation of cytokine secretion positive regulation of inflammatory response defense response to Gram-negative bacterium positive regulation of NF-kappaB transcription factor activity positive regulation of nitric-oxide synthase biosynthetic process intestinal epithelial structure maintenance positive regulation of macrophage cytokine production necroptotic process negative regulation of ERK1 and ERK2 cascade positive regulation of ERK1 and ERK2 cascade positive regulation of nucleotide-binding oligomerization domain containing 1 signaling pathway positive regulation of nucleotide-binding oligomerization domain containing 2 signaling pathway cellular response to lipopolysaccharide cellular response to lipoteichoic acid cellular response to mechanical stimulus apoptotic signaling pathway positive regulation of NLRP3 inflammasome complex assembly
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