ToxinSensor™ Chromogenic LAL Endotoxin Assay Kit: The kit utilizes a modified Limulus Amebocyte Lysate and a synthetic color producing substrate to quantitatively detect endotoxin chromogenically in a broad range (0.005–1 EU/ml).
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Under standard conditions, absorbance at 545 nm shows a linear relationship with concentrations in the range of 0.01 - 0.1 EU/ml. The above plot illustrates the absorbance (x-axis) of four standards with their corresponding endotoxin concentration (EU/ml) on the y-axis. A best-fit straight line is drawn among these points to graphically determine the endotoxin concentrations of samples based on their absorbance.Examples using following data:
|Vial No. ||Sample ||Absorbance at 545 nm ||Δ Absorbance |
|LAL reagent water (Blank) ||0.035 ||- |
|0.01 EU/ml Standard ||0.088 ||0.053 |
|0.025 EU/ml Standard ||0.133 ||0.098 |
|0.05 EU/ml Standard ||0.217 ||0.182 |
|0.1 EU/ml Standard ||0.427 ||0.392 |
If the mean absorbance of a sample is x, the endotoxin concentration of sample will be 0.2618x – 0.0012 EU/ml. All incubations were performed for 45 min. Above is only an example curve show, and the OD values of standards may be different with different assays.
ToxinSensor™ Gel Clot Endotoxin Assay Kit
: It is a qualitative test for Gram-negative bacterial endotoxin. Limulus Amebocyte Lysate as supplied is to be reconstituted with LAL Reagent Water and then mixed in equal parts with the solution being tested. After incubation, and in the presence of endotoxin, gelation occurs; in the absence of endotoxin, gelation does not occur.Note: For specimen preparation, the specimen should be certified free of Beta Glucans contaminant which may come from yeast and cellulosic materials, such as blood products.
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ToxinSensor™ Single Test Kit: is designed to be a qualitative In Vitro end-point endotoxin test for human and animal parenteral drugs, biological products, and medical devices.
Limulus Amebocyte Lysate (LAL) as supplied is to be reconstituted with sample or control directly. There is no need to reconstitute the system with LAL Reagent Water before use. After incubation for 1 hour, and in the presence of endotoxin, gelation occurs; in the absence of endotoxin, gelation does not occur.
GenScript supply a series of ToxinSensor™ Single Test Kits with different sensitivity (0.015 EU/ml, 0.03 EU/ml, 0.06 EU/ml, 0.125 EU/ml and 0.25 EU/ml).
When using the ToxinSensor™ Single Test Kit, you just need to add 200µl of sample or control to LAL, then, wait for the required incubation time to allow for gel to be formed.
Each assay should include both a positive control and a negative control (LAL Reagent Water). Following are application example:
- Sample: Protein A, 1 mg/ml in PBS, pH 7.4. It is purified from E. coli sonicate by Ni-NTA resin.
- Dilution by LAL water: 1: 200,000, 1: 400,000, 1: 800,000
- The test is performed as described above and the assay result is:
|Positive control ||Negative control ||1: 200,000 ||1: 400,000 ||1: 800,000 |
- Endotoxin concentration in this sample is: from 200,000×0.25 to 400,000×0.25, that is, from 50,000 to 100,000 EU/ml.