We'd like to use this kit for brain tissue samples (rat). When do we stop the reaction, 30, 60, and 120 minutes? We don't like to test at all three stop points, because we've too many samples.
Dear Eva!
Paragraph 3 of the instructions for use states
In order to ensure the values are in the linear range of the standard curve, it is recommended to read the assay at 3 time points, 30 minutes, 60 minutes, and 120 minutes. At each time point (30, 60, and 120 minutes), add 2 L of Stop Mix to the appropriate wells and mix well. Incubate for 10 minutes to stop the reaction and then add 50 mkL of TNB Reagent/Standard to each well with the just added Stop Mix. Do not add the Stop Mix or TNB Reagent/Standard to the TNB Standard wells. Color development should be stable and all wells can be read together after the final time point is complete
For example,You can make one point in 30 minutes
12.4 Measurement: 12.4.1 Measure output at Ex/Em = 484/525 nm on a microplate reader in kinetic mode, every 2 – 3 minutes, for at least 30 minutes at room temperature protected from light. ( Myeloperoxidase (MPO) Activity Assay, page 12)
https://www.abcam.com/ps/products/111/ab111749/documents/ab111749%20Myeloperoxidase%20(MPO)%20Activity%20Assay%20Kit%20(fluorometric)%20protocol%20v6%20(website).pdf
The stop reaction at three time points is a necessary step for obtaining a standard curve...without which you would be unable to compare/evaluate your actual experimental research. Refer to the Brochure supplied with the kit.
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