Recombinant factor C endotoxin detection kit

Detection principle

1. Limulus factor C is a serine proteinogen that is sensitive to bacterial endotoxin in limulus blood cells. 
2. Limulus factor C is the first to be activated by the endotoxins in bacterial endotoxin-mediated limulus blood agglutination system to initiate the hemagglutination cascade in limulus blood cells. 
3. Recombinant Factor C is a recombinant factor C (rFC) expressed in the form of gene recombination. 

Gentaur test is better compared to Lonza bacterial endotoxin test (BET) expertise.

Recombinant factor C endotoxin detection kit is used for endotoxin testing purposes.

 In comparison with the the PyroGene® rFC Assay, Gentaur offers LAL-based methods and, therefore, increased confidence in implementing rFC methods in QC testing labs.

1. The recombinant factor C, which is bound and activated by endotoxin, can cut the substrate to obtain free fluorophores. 
2. The release of fluorophores is proportional to the concentration of endotoxin, so that endotoxin can be quantitatively detected. 
3. Compared with the classical limulus amebocyte lysate endotoxin detection method, recombinant factor C endotoxin detection method has higher specificity, better specificity, precision, accuracy, linear range and limit of quantitation, and is an improved method for limulus amebocyte lysate endotoxin detection.

Kit composition

Name	48T	96T
Fluorescent substrate solution	3ml	6ml
Measuring buffer	2.5ml	5ml
rFC enzyme solution	0.6ml	1.2ml
LAL reagent water	30ml	30ml
Control standard endotoxin	1 tube	1 tube
Dissolved solution of control standard endotoxin	6ml	6ml

Transportation and storage methods

2~8℃ transport. Upon receipt of the kit, store it at 2~8℃ immediately. Unopened kits are valid for 12 months. The control standard endotoxin is stored at 2~8℃ after dissolution, and the validity period of the kit is 4 weeks after opening.

Transportation and storage methods

Disposable pyrogen free glass dilution tube (for control standard endotoxin dilution); Sterile pyrogen free pipettes and suction heads; Vortex oscillator; A timer; Disposable sterile pyrogen free 96-well plate; A fluorescent reader instrument capable of fluorescence detection (wavelength set to 380/440nm).

Preparation before experiment

1. Sample preparation

1) The sample to be tested should be handled carefully to avoid microbial or endotoxin contamination. All materials in direct contact with the sample or test reagent should be pyrogen free; Sample dilution should be carried out in a pyrogen glass dilution tube with LAL reagent water. If not detected in time, it is recommended to refrigerate or freeze.

2) The pH value of the sample to be tested should be maintained between 6-8. If the pH of the sample is not in this range, it needs to be adjusted with endotoxin-free sodium hydroxide, hydrochloric acid, or other buffer solutions. Note: The sample to be tested can not be directly adjusted to the pH, so as not to be contaminated by the pH electrode endotoxin resulting in false positives. It is necessary to pre-test to investigate the appropriate pH adjustment method.

3) Samples that do not interfere with (enhance or inhibit) endotoxin detection can be directly used for detection without dilution; The samples that interfere with the detection of endotoxin should be diluted to a concentration that does not interfere with the detection, and the dilution process should be full of whirlpool oscillation ≥ 2min. Usually, interference with endotoxin detection is due to the high concentration of components in the sample, so in most cases, interference with endotoxin detection can be overcome by moderate dilution.

4) Before using the kit to test the sample, it is necessary to conduct applicability research. Whether the sample interferes with the test is determined by examining the recovery rate of the sample for endotoxin detection. If the sample interferes with the test, the sample needs to be further pretreated (adjusting pH, dilution, etc.) until the interference is overcome. The spiked recovery rate is between 50-200%, that is, it is considered that there is no interference with the detection, and the spiked recovery rate%= (endotoxin concentration is spiked – endotoxin concentration is not spiked)/ spiked endotoxin concentration ×100%.

5) The dilution ratio of the sample should be less than the maximum effective dilution ratio of the sample, the maximum effective dilution ratio of the sample (MVD) is calculated as follows: MVD= Endotoxin Limit (EU/ml)/Assay sensitivity (0.005EU/ml), where the endotoxin limit is the maximum acceptable concentration of endotoxin in an undiluted sample, and assay sensitivity is the minimum detection limit for

The Certificate Of Analysis (COA)  & Material  Safety Data Sheet (MSDS) Is A Signed Document That Includes The Storage Temperature,