Intended Use
Bioendo rCR Endotoxin Test Kit (Recombinant Cascade Reagents Chromogenic Assay) is intended for use in the In Vitro quantitative detection of gram-negative bacterial endotoxins (Lipopolysaccharides) with recombinant cascade reagents chromogenic method.

Explanation of Test
The recombinant cascade reagents chromogenic endotoxin assay is a quantitative assay for the detection of gram-negative bacterial endotoxins. The assay is conducted by mixing Recombinant Cascade Reagents with test sample followed by recording the mixture’s absorbance at 405nm at 37oC. The velocity of the absorbance increase is related to the endotoxin concentration of the test sample. A fast absorbance increase indicates high endotoxin concentration, whereas a slow one indicates low endotoxin concentration. Endotoxin concentration is determined based on the fastness of the absorbance increase.

Principles
Bioendo rCR Endotoxin Test Kit (Recombinant Cascade Reagents Chromogenic Assay) uses gene recombinant technology to express Factor C pathway serine zymogen proteases of Tachypleus amebocyte in eukaryotic cells. The cascade reagents include Factor C which can be activated by bacterial endotoxins, Factor B and proclotting enzyme. Endotoxins bind to and convert Factor C to its active form, which subsequently activates Factor B, which in turn activates the proclotting enzyme. The activated proclotting enzyme then cleaves colorless substrate, releasing a yellow-colored pNA. The released pNA can be measured photometrically at 405 nm. With the formation of pNA, the absorbance at 405nm is increased. The velocity of the absorbance increase is positively correlated with the endotoxin concentration. That is, the time needed to reach a certain absorbance increase (onset OD), termed onset time, is negatively correlated with the endotoxin concentration. Based on which, the endotoxin concentration of test sample can be quantified.

Storage Conditions
Bioendo rCR Endotoxin Test Kit (Recombinant Cascade Reagents Chromogenic Assay) is stored at 2-8°C.

Reagents Supplied
1. Recombinant Cascade Reagents, Catalog No. RCR0428S or RCR0428, Blue-Labeled Vial
2. Reconstitution Buffer, Catalog No. RCRRB28, Green-Labeled Vial
3. Control Standard Endotoxin (CSE), Catalog No. CSE10V, Red-Labeled Vial
4. Water for BET, Catalog No. TRW50, Grey-Labeled Vial