Thermo Fisher Scientific CD284 (TLR4) Monoclonal Antibody (HTA125), Super Bright 645, eBioscience

Thermo Fisher Scientific CD284 (TLR4) Monoclonal Antibody (HTA125), Super Bright 645, eBioscience

Applications and Tested Dilution

Product Specifications

Available 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 600
• Request custom conjugation available

Product Specific Information

The HTA125 monoclonal antibody reacts with human Toll-like receptor 4 (TLR4). TLR family members are type I transmembrane proteins with leucine-rich extracellular domains and IL-1 receptor-like cytoplasmic domains. TLR4 associates with MD-2 and CD14, forming a complex responsible for LPS recognition and signaling. TLR4 is expressed by peripheral blood monocytes.

HTA125 immunoprecipitates human TLR4 (~100 kDa) from transfected cells. Surface expression of TLRs, particularly TLR1 and TLR4, is low on monocytes and even lower on granulocytes and immature dendritic cells. Expression levels vary among donors.

For optimal staining of TLR4, use the lysed whole blood protocol rather than Ficoll-gradient prepared cells, as the latter reduces staining intensity.

Applications Reported: Flow cytometric analysis
Applications Tested: Pre-diluted and tested by flow cytometry using human TLR4-transfected cells. Recommended use: 5 µL (0.5 µg)/test in 100 µL final volume.

Fluorochrome Information:
Super Bright 645 is a tandem dye excited by the violet laser (405 nm) and emits at 645 nm. Recommended filter: 660/20 bandpass. Ensure instrument compatibility.

Staining Panel Recommendation:
When using multiple Super Bright dye-conjugated antibodies, use Super Bright Complete Staining Buffer (Product # SB-4401-75) to minimize non-specific polymer interactions.

Handling and Storage:

• Light sensitive: protect vial and stained samples from light
• Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days at 4°C in the dark
• Excitation: 405 nm
• Emission: 645 nm
• Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.

Target Information

TLR4 is a member of the Toll-like receptor family involved in pathogen recognition and activation of innate immunity. It is expressed mainly in placenta and myelomonocytic leukocytes. TLR4 forms a complex with CD14 and LY96 to mediate LPS-induced signaling in response to gram-negative bacteria. Mutations in TLR4 affect LPS responsiveness. TLR4 expression is found in monocytes, a small population of B-cells, and placenta. Studies with TLR4-deficient mice confirm its role in LPS recognition and susceptibility to Gram-negative sepsis.

For Research Use Only. Not for use in diagnostic procedures.

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