China Suppliers: Sustainable rFC Endotoxin Assay Kit for Reliable BET from a Certified Factory
Technical Information on Recombinant Factor C (rFC) Fluorometric Assay for Bacterial Endotoxin Detection
1. Assay Principle
Endotoxin LPS specifically binds to the LPS-recognition domain of rFC, triggering a conformational change that activates rFC’s serine protease activity. The intensity of AMC fluorescence is directly proportional to the endotoxin concentration in the sample, measurable via a fluorescence microplate reader.
2. Key Technical Parameters
Detects endotoxins at concentrations as low as 0.005 EU/mL, matching or exceeding traditional Limulus Amebocyte Lysate (LAL) assays.
Typically covers 0.005–50 EU/mL, enabling quantitative analysis across a broad concentration spectrum.
Exhibits high specificity for LPS; it is not activated by (1→3)-β-D-glucans (a common interference in LAL gel-clot assays), reducing false-positive results.
Requires controlled temperature (37±1°C) and incubation time (60–120 minutes), with fluorescence measured at excitation/emission wavelengths of ~360 nm/~460 nm.
3. Workflow Overview
Prepare samples (e.g., pharmaceuticals, medical devices, biological products) via dilution or extraction to remove matrix interference.
Mix rFC reagent, fluorogenic substrate, and sample in a microplate well.
Incubate at 37°C to allow LPS-rFC binding, activation, and substrate cleavage.
Measure fluorescence intensity; calculate endotoxin concentration using a standard curve generated with known endotoxin standards (e.g., Escherichia coli O55:B5 LPS).
This assay combines sustainability (no horseshoe crab harvesting), reliability (low interference, high reproducibility), and efficiency, making it ideal for quality control in pharmaceutical and biotech industries.