Recombinant Factor C Endotoxin Detection Kit HYI131
Recombinant factor C endotoxin detection kit can replace the traditional limulus amebocyte lysate (LAL) for human and animal injection drugs (such as chemicals, radiopharmaceuticals, antibiotics, biological products, etc.) and medical devices (such as dialysate, implantable devices, etc.) raw and auxiliary materials, intermediate products, released products endotoxin detection. This kit is a highly sensitive, highly specific, stable and sustainably available alternative to limulus amebocyte lysate that does not depend on animal-derived components. It can measure endotoxin concentrations in the range of 0.005-5 EU /mL.
Detection principle
Limulus factor C is a serine proteinogen sensitive to bacterial endotoxin in limulus blood cells. Limulus factor C is the first to be activated by endotoxin in bacterial endotoxin-mediated limulus blood agglutination system to initiate the hemagglutination cascade in limulus blood cells. Recombinant Factor C is a recombinant factor C (rFC) expressed in the form of gene recombination. The recombinant factor C, which is bound and activated by endotoxin, can cut the substrate to obtain free fluorophores. The release of fluorophores is proportional to the concentration of endotoxin, so that endotoxin can be quantitatively detected. 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.
Components
|
Reagent |
48T |
96T |
|
Fluorescent substrate solution |
3 mL |
6 mL |
|
Measuring buffer |
2.5 mL |
5 mL |
|
rFC enzyme solution |
0.6 mL |
1.2 mL |
|
LAL reagent water |
30 mL |
30 mL |
|
Control standard endotoxin |
1 vial |
2 vials |
|
96-well non-pyrogenic detachable reaction plate |
1 Pc |
1 Pc |
Note: Control standard endotoxin is freeze-dried powder. See label for titer and reconstitution volume.
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 is up to 8 weeks.
Operation brief
The assay was performed in a 96-well plate, and the fluorescence values were measured at an excitation/emission wavelength of 380/440nm at zero hour after sample addition and one hour after incubation at 37°C ±1°C. The net fluorescence difference is obtained by calibrating the fluorescence reading difference (ΔRFU) between the control standard endotoxin and the sample using the fluorescence difference (ΔRFU) of the negative control. The logarithm of the net fluorescence difference is proportional to the logarithm of the endotoxin concentration and is linear over the range 0.005 to 5.0EU/mL. The endotoxin concentration in the sample can be calculated according to the standard curve.
Required consumables and equipment
Disposable pyrogen free glass dilution tube (for control standard endotoxin dilution); Sterile pyrogen free pipettes and suction heads; Vortex oscillator; A timer; A fluorescent reader instrument capable of fluorescence detection (wavelength set to 380nm/440nm).