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Poor signal to noise ratio despite good incorporation
Low signal can be caused by inappropriate hybridization conditions (too stringent buffer or excessive temperature) or when the hybridization time…
Tue, Jul 18, 2023
How stable is the Cyto-ID® Green autophagy dye?
Fluorescent dyes are subject to photobleaching, but this dye is more robust than dyes like fluorescein. Standard handling precautions should…
Tue, Jul 18, 2023
Are the cells fixable after staining with the autophagy kit?
Yes, a protocol for fixing cells is provided in the product manual. Permeabilization of the cellular membrane with detergents, however,…
Tue, Jul 18, 2023
*General Information
The solubility of small molecule products, especially those that are hydrophobic can sometimes be a limiting factor in experimental design.…
Tue, Jul 18, 2023
Little or no apoptosis observed or detected
None or few apoptotic cells present in sample >> Use the Apoptosis Inducer, provided in the kit, as a positive…
Tue, Jul 18, 2023
Precipitate is seen in the 10X assay buffer
Precipitate forms at low temperatures. Allow solution to warm to room temperature or 37°C.
Tue, Jul 18, 2023
Good DLRs in the CGH analytics QC report, but some oligos show whiskering or significant deviation from the expected log2 ratio
We suggest keeping the Cot-1 DNA at 50 µg regardless of the hybridization volume.
Tue, Jul 18, 2023
Bright yellow array with reduced magnitude of expected changes
This can be due to insufficient Cot-1 DNA, blocking agents or hybridization and washing conditions that are not stringent enough.
Tue, Jul 18, 2023
What is the detailed nature of the Cyto-ID® autophagy dye and how does the assay work?
The precise chemical structure of the dye is proprietary. The probe is a cationic amphiphilic tracer (CAT) dye that rapidly…
Tue, Jul 18, 2023
What are the advantages of the Cyto-ID® autophagy dye relative to monodansyl cadaverine (MDC)?
MDC requires 365 nm UV illumination and thus, is not compatible with 488 nm excitation sources commonly implemented in flow…
Tue, Jul 18, 2023