Applications: Suitable for kinetic and endpoint assays using Horseradish peroxidase (HRP) labeled probes.
Protocol Overview: 1. Complete all required incubations with antibodies, probes and HRP labeled reagents.
2. Wash plates, at least 4 times, with phosphate buffered saline or Tris-buffered saline containing 0.1% Tween-20.
3. After the final wash, shake and blot all residual buffers from plate wells.
4. Add 100 µl of ABTS solution to each well and incubate at room temperature for 30 minutes. Readings at 405nm can be taken at predetermined intervals if a kinetic assay is desired.
5. After 30 minutes, 100 µl of acid can be added as stop solution and read at 405nm. The colour will be stable for at least 1 hour if 0.625M oxalic acid is used.
Shipping Conditions: The reaction time will depend upon the activity of the HRP probe. If color develops too briskly, zero order kinetics will not prevail. Dilution of a probe, antibody, HRP labeled reagent may be required. In addition, time, reagent volume and temperature variations may require standardization by the user.
Storage Condition: Store at room temperature (17-28ºC). Storage at refrigerator temperatures (2-4ºC) will not inhibit product performance but warm the product to assay temperatures prior to use. Protect from exposure to direct sunlight and discard the solution if it becomes yellow and turbid.