Detection range: 96T
0.03Eu/L – 0.8Eu/L
Kit composition
1 | 30 times concentrated washing solution | 20ml × 1 bottle | 7 | Stop solution | 6ml × 1 bottle |
2 | Enzyme standard reagent | 6ml × 1 bottle | 8 | Standard product (1.6Eu/L) | 0.5ml × 1 bottle |
3 | Enzyme label coated plate | 12 holes × 8 | 9 | Standard dilution | 1.5ml × 1 bottle |
4 | Sample diluent | 6ml × 1 bottle | 10 | Instruction manual | 1 copy |
5 | Developer A solution | 6ml × 1 bottle | 11 | Sealing film | 2 sheets |
6 | Developer B solution | 6ml × 1 bottle | 12 | sealed bag | 1 |
E. coli endotoxin enzyme-linked immunoassay kit procedure
1. Dilution of standard: This kit provides one original standard, which can be diluted in a small tube according to the following chart.
0.8Eu/L | Standard No. 5 | Add 150 μl of standard dilution to 150 μl of the original standard |
0.4Eu/L | Standard No. 4 | Add 150 μl of standard dilution to 150 μl of standard #5 |
0.2Eu/L | Standard No. 3 | Add 150 μl of standard dilution to 150 μl of standard #4 |
0.1Eu/L | Standard 2 | Add 150 μl of standard dilution to 150 μl of Standard #3 |
0.05Eu/L | Standard No. 1 | 150 μl of Standard 2 is added to 150 μl of standard dilution |
2. Adding samples: set blank holes separately (the blank control wells are not added with the sample and the enzyme standard reagent, the other steps are the same), the standard holes, and the sample holes to be tested. Accurately load 50 μl of the standard on the enzyme-labeled plate, add 40 μl of the sample dilution to the well to be tested, and then add 10 μl of the sample to be tested (the final dilution of the sample is 5 times). Add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, and shake gently to mix.
3. Incubation: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes.
4. Liquor: 30 times concentrated washing solution diluted with distilled water 30 times and used
5. Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing solution, let stand for 30 seconds, then discard it, repeat 5 times, and pat dry.
6. Add enzyme: Add 50 μl of enzyme labeling reagent to each well, except for blank wells.
7. Incubation: The operation is the same as 3.
8. Wash: Operate the same as 5.
9. Color development: add 50 μl of color developer A, and then add 50 μl of color developer B, gently shake and mix, and color for 15 minutes at 37 °C.
10. Termination: 50 μl of stop solution was added to each well to stop the reaction (the blue color turned yellow).
11. Measurement: The absorbance (OD value) of each well was measured in sequence with a blank air conditioner of zero and a wavelength of 450 nm. The measurement should be carried out within 15 minutes after the addition of the stop solution.
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