ToxinSensorTM 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).
Note:
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Technical Advantages:
Under the standard conditions, the absorbance at 545 nm shows a linear relationship with the concentration in range of both 0.01 - 0.1 EU/ml and 0.1 - 1 EU/ml. Plot the absorbance for the four standards on the x-axis, the corresponding endotoxin concentration in EU/ml on the y-axis. Draw a best-fit straight line among these points and determine endotoxin concentrations of samples graphically.
Examples using following data:
| Tube No. |
Sample |
Absorbance at 545 nm |
Δ Absorbance |
| 1 |
LAL reagent water (Blank) |
0.035 |
- |
| 2 |
0.01 EU/ml Standard |
0.088 |
0.053 |
| 3 |
0.025 EU/ml Standard |
0.133 |
0.098 |
| 4 |
0.025 EU/ml Standard |
0.217 |
0.098 |
| 5 |
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.
ToxinSensorTM 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.
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Technical Advantages:
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.
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