1. Dilution of standard: This kit provides one original standard, which can be diluted in a small tube according to the following chart.
2. Loading: 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. Dosing: 20 times concentrated washing solution diluted with distilled water 20 times and used
5. Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing liquid, let stand for 30 seconds, then discard it, repeat 5 times, and pat dry.
6. Add enzyme: 50 μl of enzyme labeling reagent was added to each well, except for blank wells.
7. Incubation: The operation is the same as 3.
8. Washing: The operation is the same as 5.
9. Color development: Add 50 μl of the developer to each well, then add 50 μl of the developer, gently shake and mix, and color at 37 ° C for 15 minutes.
10. Termination: 50 μl of stop solution was added to each well to terminate the reaction (when the blue color turned yellow).
11. Measurement: The absorbance (OD value) of each well was measured sequentially 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.
Storage conditions and expiration date
1. The kit is stored at: 2-8 °C.
2. Rat Bcl-2 Related X Protein (BAX) ELISA Kit Validity: 6 months
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