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Knowledge Base
Edge Effects
Where was the plate incubated? Often times the conditions for ambient incubations can be less than ideal. If there is…
Tue, Jul 18, 2023
Poor Standard Curve
What was used as the standard diluent? Diluents other than the supplied assay buffer may contain interfering substances that can…
Tue, Jul 18, 2023
Poor Precision
Were the wells washed properly? All wells receive the same treatment during the wash step. If some are washed less…
Tue, Jul 18, 2023
Drift
Were reagents brought to room temperature prior to use? If the reagents are not at a constant temperature prior to…
Tue, Jul 18, 2023
Weak Color Development
Was substrate added at the correct point in the assay? See the assay procedure provided in the instruction manual. Were…
Tue, Jul 18, 2023
High Background
How was the plate washed? It is important that the plate is washed thoroughly. If plate washing is troublesome, a…
Tue, Jul 18, 2023
How long can the Cyto-ID® green detection reagent be left incubating with live cells (mins/hours/days)?
We do not recommend to incubate cells with Cyto-ID® green detection reagent for longer that 30 minutes at 37°C. In…
Tue, Jul 18, 2023
How soon after incubation with Cyto-ID® autophagy dye would the samples need to be analyzed by flow cytometry? The user manual recommends following 30 minutes incubation. Is it advisable to temporarily store samples on ice before they are analyzed?
One can store the samples on ice for a short period of time to extend the period of optimal staining.…
Tue, Jul 18, 2023
For the ROS/RNS detection kit, ENZ-51001, can samples be stored after the final wash buffer step in the protocol, and do the microscopy at a later date?
No, this is a vital stain. Live cells must be read immediately per the manual.
Tue, Jul 18, 2023
How long can the fluorescence signal be detected in live cells?
Typically, cells are induced to undergo autophagy and then stained just prior to analysis. Induction can be as little as…
Tue, Jul 18, 2023