Anti-TLR4 mAbs

Cat. No.

MABL-3113

Name

Anti-TLR4 mAbs

Clone No.

AFD- NI-0101

From

Recombinant Antibody

Isotype

Engineer antibody

Application

crystallization, FACS, functional assays,   inhibition, SPR, ELISA

Species Reactivity

Human

Basic Information

Specificity

This antibody is specific for human TLR4 and binds the LRR 9 domain (residues 328-YN-329, 349-KLK-351); it engages both   FcγRI and FcγRIIA, but not FcγRIII.

Alternative Name

hToll; CD284; Toll-like receptor 4; hu 15C1;   hu15C1

UniProt

O00206

Immunogen

The original antibody was generated through   a humanization process involving CDR grafting of the original mouse anti-TLR4   clone 15C1 onto selected human germline sequences and performing framework   optimization.

Application Notes

The original antibody (human IgG1) was   generated and used for SPR, FACS, and various functional assays. SPR   determined the binding affinity of Hu 15C1 with Fcγ receptors and TLR4,   showing strong binding to FcγRI (47.3 ± 1.3 nM) and FcRn (1.82 ± 0.09 nM) and   moderate binding to FcγRIIA 131R (0.151 ± 0.015 µM), FcγRIIA 131H (0.311 ±   0.037 µM), and FcγRIIB (0.870 ± 0.038 µM). FACS assays evaluated Hu 15C1   binding to CHO cells expressing human TLR4 and MD-2. Functional assays   included human macrophage and U937 cell assays to measure IL-6 production,   confocal microscopy, and FRET analysis to visualize TLR4 and Fcγ receptor   interactions, and LPS lung instillation in mice to assess cytokine and   antibody levels in bronchoalveolar lavage fluid.

Antibody First   Published

 Shang et al., Selective antibody   intervention of Toll-like receptor 4 activation through Fc γ receptor   tethering. J Biol Chem. 2014 May 30;289(22):15309-18. PMID:24737331

Note on publication

The original publication describes the   humanization and characterization of this antibody clone and a novel   mechanism where anti-TLR4 antibodies are designed to tether both TLR4 and Fcγ   receptors.

COA Information （For reference only, actual COA shall prevail）

Size

100 μg Purified   antibody.

Concentration

1 mg/ml.

Purification

Protein A affinity   purified

Buffer

PBS with 0.02% Proclin   300.

Concentration

1 mg/ml.

Storage   Recommendation

Store at 4⁰C for up to 3   months. For longer storage, aliquot and store at - 20⁰C.