Thermo Fisher Scientific CD284 (TLR4) Monoclonal Antibody (HTA125), Super Bright 600, eBioscience

Applications

Tested Dilution

Publications

Western Blot (WB)

-

View 1 publication 1 publication

Immunohistochemistry (IHC)

-

View 1 publication 1 publication

Flow Cytometry (Flow)

5 µL (0.5 µg)/test

View 6 publications 6 publications

Neutralization (Neu)

-

View 4 publications 4 publications

Product Specifications

Published species

Not Applicable

Host/Isotype

Mouse / IgG2a, kappa

Recommended Isotype Control

Mouse IgG2a kappa Isotype Control (eBM2a), Super Bright™ 600, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

HTA125

Conjugate

Super Bright™ 600 Super Bright™ 600 Super Bright™ 600

View additional formats

• Unconjugated
• Alexa Fluor 488
• Alexa Fluor 700
• APC
• APC-eFluor 780
• Biotin
• eFluor 450
• Functional Grade
• PE
• PE-Cyanine7
• PerCP-eFluor 710
• Super Bright 436
• Super Bright 645
• Request custom conjugation

Excitation/Emission Max

414/601 nm View spectra

Form

Liquid

Concentration

5 µL/Test

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Wet ice

RRID

AB_2734948

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 BestProtocols®) 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.

Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer ([Product # SB-4401-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

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 (](https://www.thermofisher.com/order/catalog/product/SB-4401-42)[Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (](https://www.thermofisher.com/order/catalog/product/00-8222-49)[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: 405 nm; Emission: 600 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.](https://www.thermofisher.com/order/catalog/product/00-5333-57)

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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.

](https://www.thermofisher.com/order/catalog/product/00-5333-57)