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Cellular Analysis
Fluorescence does not increase after incubation with inhibitor.
Cells do not express any MDR1, MRP1/2 or BCRP. Use a positive control cell type expressing corresponding ABC transporter.
Tue, Jul 18, 2023
There are differences in MFI (mean fluorescence intensity) values between cell lines.
Detection reagent accumulation may be influenced by cell size, endogenous esterase activity, etc. Using the inhibitors and calculating the MAF…
Tue, Jul 18, 2023
Inconsistent fluorescence shift using the same cell (irreproducible results).
Inadequate incubation condition. Always use a water bath (not incubator) and ensure temperature of water bath is 37°C.
Tue, Jul 18, 2023
Do I need a luminometer with double injectors for this assay?
Yes. A single injector will not provide good results. You cannot pre-mix the arachidonic acid and substrate to make a…
Tue, Jul 18, 2023
Can I use cell culture samples in the COX Activity assay?
Cell culture samples are compatible with this kit. Since this is an activity assay rather than a quantitative immunoassay, there…
Tue, Jul 18, 2023
In the sPLA2 activity kit (ADI-907-002), how is a Unit defined?
One unit of phospholipase activity is the amount of enzyme needed to hydrolyze 1.0 µmole of L-a-phosphatidylcholine to L-a-lysophosphatidylcholine and…
Tue, Jul 18, 2023
DNA histograms are of poor overall quality.
The key elements in obtaining a histogram of high quality are good sample preparation, correct cell-to dye ratio and proper…
Tue, Jul 18, 2023
No decrease of the fluorescent signal after using a specific inhibitor
Inhibitor concentration is too low or too high. Very low doses of inhibitor may not affect ROS production by inducer.…
Tue, Jul 18, 2023
Low or no fluorescent signal in positive control
Species of interest may react with each other, thus attenuating the expected signal. ROS is a complex interacting system. Blocking…
Tue, Jul 18, 2023
Can I use the COX activity kit to distinguish between COX-1 and -2?
The COX Activity kits measure activity alone, distinguishing between COX-1 or -2. In order to differentiate between the two enzymes,…
Tue, Jul 18, 2023