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Cells do not exhibit fluorescence after incubation with the detection reagent.
Cell viability is low. Cells should be in log growth phase. Cell samples cannot be kept longer than 6 hours…
Tue, Jul 18, 2023
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
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
More than one aggregation peak is observed.
A protein might have multiple domains that start aggregation at different temperatures. For most samples, any aggregation is an indication…
Tue, Jul 18, 2023
Insulin does not go into solution.
Insulin was not reconstituted in deionized water prior to dilution. Resuspend Insulin in deionized water before dilution into PBE Buffer.
Tue, Jul 18, 2023