Rat TLR4 (Toll-Like Receptor 4) CLIA Kit (RTES00561)

Key Features & Specifications

This chemiluminescent immunoassay provides quantitative detection of Toll-Like Receptor 4 across a broad dynamic range, leveraging an enhanced luminescent substrate that delivers higher sensitivity than conventional colorimetric ELISA formats. The assay utilizes a sandwich immunoassay configuration in which captured TLR4 is bound by a biotinylated detection antibody and visualized using streptavidin conjugated to horseradish peroxidase.

• Format: 96-well microplate, pre-coated and ready to use
• Sample volume: typically 50-100 ul per well
• Assay time: approximately 3.5 hours from sample loading to signal readout
• Detection principle: chemiluminescence, read on a luminometer or suitable plate reader
• Calibration: recombinant rat TLR4 standards included for generating a standard curve
• Sample compatibility: serum, plasma (EDTA, heparin), tissue homogenates, cell culture supernatants

Workflow & Data Interpretation

Briefly, add standards and samples to coated wells, incubate, wash, add the biotinylated antibody, incubate again, wash, add streptavidin-HRP, wash, and finally add the chemiluminescent substrate. Signal intensity is proportional to TLR4 concentration and is interpolated against the standard curve. Suggested dilution ranges for unknown samples are typically 1:2 to 1:10, although optimization may be required for specific matrices.

Storage & Stability

Store unopened components at 2-8 degrees C. Reconstituted standards and working detector solutions should be stored refrigerated and used within the timeframe indicated in the product insert. Avoid repeated freeze-thaw cycles of reconstituted standards. Protect substrate and streptavidin-HRP from prolonged light exposure to preserve activity. For research use only.

Research Applications

TLR4 is widely studied in the context of innate immunity, endotoxin recognition, sepsis, neuroinflammation, metabolic inflammation, and host-pathogen interactions. This kit, part of the Assay Genie portfolio of inflammation and immunology reagents, supports biomarker profiling in experimental rat models investigating LPS challenge, MyD88-dependent and TRIF-dependent signaling pathways, and downstream NF-kB activation.