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Catalog Number | Quantity |
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Q32892 | 160 reactions |
The Invitrogen Quant-iT Endotoxin Detection Assay Kit is a sensitive and easy-to-use fluorescent endpoint assay for use on fluorescence microplate readers that uses amebocyte lysates to quantify endotoxin in protein, peptide, antibody, or nucleic acid samples. Amebocyte lysates are widely used as a simple and sensitive assay for the detection of endotoxin lipopolysaccharide (LPS), the membrane component of gram-negative bacteria.
Features of the Quant-iT Endotoxin Detection Assay Kit include:
• Highly sensitive with a broad range—detect as little as 0.01 EU/mL to 10.0 EU/mL
• Flexible—suitable for wide range of samples, including DNA, RNA, plasmids, antibodies, and more
• Easy to use—the simple workflow requires one incubation prior to measurement
Endotoxin levels in the samples are accurately determined using the included endotoxin standard of known concentration that is derived from E. coli strain O111: B4. Determining endotoxin levels is important to assess the efficiency of endotoxin removal methods and prevent endotoxic shock, inflammation, and/or sepsis in tissue culture cells and animals injected with endotoxin-contaminated proteins.
When endotoxin encounters the amebocyte lysate, a series of enzymatic reactions results in the activation of Factor C, Factor B, and pro-clotting enzyme. The activated enzyme catalyzes a cleavage event in the substrate to produce a strong fluorescent signal. After stopping the reaction, the resulting signal is measured on a fluorescence microplate reader (such as Varioskan LUX). The correlation between fluorescent signal is proportional to the endotoxin concentration in the sample.
Notes
• Compatible with Pierce Endotoxin-Free Plate Strips (Cat. No. A43888)
The US Pharmacopeia recommends performing this analysis using log transformed concentration and RFU data and then performing a linear regression fit to verify that the correlation coefficient, r, is greater than or equal to 0.980.
The entire kit can be stored at 2-8 degrees C for 6 months. Reconstituted Endotoxin standard solutions are stable for 4 weeks at 2-8 degrees C and reconstituted dye can be stored for 4 weeks at 2-8 degrees C. Reconstituted amebocyte lysate solution is stable for 1 week at -20 degrees C or colder if frozen immediately after reconstitution.
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