Q: What samples types are suitable for this assay?
A: Our AOPP Assay can be used for any samples containing AOPP above the kit detection limit, such as plasma, serum, tissue and cell lysates that have been stored at -80ºC for no more than one year.
Q: How do I prepare lysates for the AOPP Assay?
A: We recommend lysing cells or tissues in 1X PBS containing proteinase inhibitors by sonication or homogenization, centrifuge at 12000 xg for 10 min, and harvest the supernatant as the lysate. We only recommend using PBS for preparing lysates because we don’t know if lysis buffers will damage the AOPPs. The protein concentration should be determined with a protein assay, such as BCA or Bradford. We don’t recommend a specific protein concentration for this assay because it will be sample dependent, however it is suggested that a sample titration is performed prior to running the experiment to determine any necessary dilutions so that samples fall within the standard curve.
Q: How is the AOPP concentration of samples expressed?
A: The concentration of sample AOPP is expressed in chloramine units (µM) or µM chloramine equivalent units.