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Catalog # HM2068

TLR4, Human, mAb HTA125


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Toll-like receptors (TLRs) are highly conserved from Drosophila to humans and share structural and functional similarities. TLRs constitute of a family of pattern recognition receptors (PRRs) that mediate cellular responses to a large variety of pathogens (viruses, bacteria, and parasites) by specific recognition of so-called ‘pathogen-associated molecular patterns’. Activation of TLRs, a family of at least 11 different members that function either as homo- or heterodimers, leads to activation of NFκB-dependent and IFN-regulatory factor-dependent signaling pathways. TLRs have a central role in innate immunity and are also required for the development of an adaptive immune response. TLRs are expressed by various cells of the immune system, such as macrophages and dendritic cells. TLRs are class I receptors, with a single α-helix that spans the cell membrane. They recognize and respond to molecules derived from bacterial, viral and fungal pathogens, such as lipopolysaccharide (LPS) from the outer membrane of Gram negative bacteria, peptidoglycan fragments from bacterial cell walls and single-stranded and double-stranded RNA from viruses. Toll-like receptor 4 (TLR4; CD284) has been identified, next to MD-2 and CD14, as a receptor that is central to the innate immune response to LPS of Gram-negative bacteria. TLR4 is unique among TLRs in its ability to activate two distinct signaling pathways; one pathway is activated by the adaptors TIRAP (Toll/interleukin-1- receptor (TIR)-domain-containing adaptor protein) and MyD88, which leads to the induction of pro‑inflammatory cytokines. The second pathway is activated by the adaptors TRIF (TIR-domaincontaining adaptor protein inducing interferon‑β) and TRAM (TRIFrelated adaptor molecule), which leads to the induction of type I interferons.
The monoclonal antibody HTA125 is a TLR4 function-blocking antibody. HTA125 recognizes preferentially human TLR4 that is associated with MD-2.

Flow cytometry, Functional studies, Immuno assays, Immuno fluorescence, Immuno precipitation, Western blot

Application Notes
FC: 300000 cells/50 µl were stained with 2  µg antibody for 30 minutes at 4 °C
FS: In cell culture 10  µg/ml
IF: Oregon green labeled HTA125 was used in FRAP measurements
IP: HTA125 (4 mg/ml) coupled to Sepharose 4FF beads was added to cell lysate and incubated for 2 hours at 4 °C
W: 20 mg protein was analyzed on SDS-PAGE and transferred to nitrocellulose. Blot was blocked with TBS/5% dry milk/0.1% tween-20

For flow cytometry and immunohistology dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For neutralization of biological activity in vitro dilutions have to be made according to the amounts of TLR4 to be inactivated. Before use in biological assays, the product must be filter sterilized and depending on the concentration to be used dialyzed against culture medium to remove the sodium azide added. Please inquire for availability of azide free solutions.

Positive Control

Product type
Monoclonal antibodies
100 µg, 20 µg
0.2 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.02% sodium azide and 0.1% bovine serum albumin.
BALB/c mice were immunized with the Ba/F3 line expressing TLR4.
Mouse IgG2a
Cross reactivity
Canine – Yes, Monkey – Yes
TLR4, CD284, ARMD10
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for one year.
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Infectious diseases

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