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Invitrogen
CD284 (TLR4) Monoclonal Antibody (HTA125), PerCP-eFluor™ 710, eBioscience™
FIGURE: 1 / 25
CD284 (TLR4) Antibody (46-9917-42) in Flow
TLR4/MD2-transfected cells were stained with Mouse IgG2a kappa Isotype Control, PerCP-eFluor 710 (Product # 46-4724-82) (blue histogram) or CD284 (TLR4) Monoclonal Antibody, PerCP-eFluor 710 (purple histogram). Viable cells were used for analysis, as determined by Fixable Viability Dye eFluor 450 (Product # 65-0863-18).
Product Details
46-9917-42
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Human
Published species
Not Applicable
Host/Isotype
Mouse
/ IgG2a, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
HTA125
Conjugate
Excitation/Emission Max
482/708 nm
View 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
Gene Aliases: ARMD10; CD284; TLR-4; TLR4; TOLL
UniProt ID: (Human) O00206
Entrez Gene ID: (Human) 7099
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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