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Cellular Analysis
Can I run cell lysates with the Nitric Oxide detection kits?
Yes. First remove the media from the cells. Pellet them and wash with PBS before lysing the cells in the…
Tue, Jul 18, 2023
My cell culture samples are in DMEM. Will this interfere with the Nitric Oxide assay?
Yes. The nitrate salt used in this particular media will cause interference. It is important to check the published media…
Tue, Jul 18, 2023
How do I know whether to use the Nitric Oxide (NO<sub>2-</sub>/NO<sub>3-</sub>) detection kit or the Nitric Oxide (total) detection kit?
The regular kit measures both major metabolic products of NO — nitrate and nitrite. The Total kit measures these metabolites…
Tue, Jul 18, 2023
Can I measure tissue NO with the Nitric Oxide detection kits?
Yes. Samples can be homogenized in the provided Reaction Buffer and then centrifuged. The supernatant should then be filtered through…
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
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
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
What is actually being stained in the nucleolus by the Nucleolar-ID™ Green Detection Reagent?
We use an RNA binding dye and this has been validated. However, in the context of live cells we cannot…
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