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Autophagy Detection
Is Autophagy Detection reagent in ENZ-51031 and ENZ-KIT175 compatible with anti-fluorescence quenching agent and fluoromount-G?
The CYTO-ID® Autophagy Detection reagent in ENZ-51031 and ENZ-KIT175 kits is compatible with anti-fluorescence quenching agent and Fluoromount-G.
Tue, Jul 18, 2023
Can Cyto-ID® Green autophagy detection reagent from Cyto-ID® Autophagy detection kit, ENZ-51031, be used in combination with ProteoStat® Detection Reagent in ProteoStat® Protein aggregation assay, ENZ-51023?
Since there is no spectral overlap between Cyto-ID® Green Autophagy Detection Reagent and ProteoStat® Detection Reagent, they can be used…
Tue, Jul 18, 2023
Blue nuclear counterstain signal is dramatically reduced in treated cells.
If the blue nuclear signal decreases by more than 30%, the testing compound is generally cytotoxic. Optimize the concentration of…
Tue, Jul 18, 2023
After incubation with the dual detection reagent at 37° C for 30 min, the cells were not healthy and rounding up. Some cells were also detached from the plate. No specific staining on positive control was observed vs. DMSO controls.
For fragile cells, the following modification to the standard protocol are suggested: Use PBS/5% FBS as an assay buffer or…
Tue, Jul 18, 2023
Is it possible to detect autophagy by fluorescence microscopy using 3D culture systems?
We have not tested the dye in 3D culture systems, but the small cell clusters are probably dye-accessible. For live…
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
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
What are the advantages of the Cyto-ID® Autophagy Detection kit relative to GFP-LC3 transfected cells?
The most significant advantage for Cyto-ID® Autophagy Detection kit is that it's a no-transfection assay. Transfections are transient and there…
Tue, Jul 18, 2023
Can the Cyto-ID® green detection dye be used in conjunction with another dye to differentiate between endosomes and amphisomes?
The Cyto-ID® Red long-term tracer dye, ENZ-51037- inserts into the plasma membrane and later is internalized by endocytosis. Cyto-ID® Red…
Tue, Jul 18, 2023