The effect of aptamers on cell viability was tested using the PrestoBlue assay (Sigma-Aldrich Ltd, Butterworth, UK). Briefly, 2000 cells/ well of U87MG, 1321N1 and SVGp12 cells were seeded in 96-well plates and incubated for 24 h. Cells were treated with four different concentrations (20 nM, 100 nM, 500 nM and 1000 nM) of biotinylated DNA aptamers SA43 or SA44 for 24, 48 and 72 hours. Cisplatin diluted in media (10 μM) was used as a positive control. To evaluate the cell viability following 24, 48 and 72 hours treatment, 10 μl of PrestoBlue was added to each well and incubated for 1 hour at 37°C. The assay was read using a TECAN genios pro multifunctional microplate reader (Tecan, Austria; software version 4.53) with excitation 535 nm and emission 612 nm. Cell viability was expressed in percentage relative to control untreated cells grown in media.
Prestoblue assay
Manufactured by Merck Group
Sourced in United Kingdom
PrestoBlue is a cell viability reagent that measures the metabolic activity of cells. It contains a cell-permeable, non-toxic, and water-soluble compound that changes color in response to chemical reduction of the growth medium resulting from cell metabolism. This change in color can be quantified using a spectrophotometer or a fluorescence plate reader, providing a measure of cell viability and proliferation.
Automatically generated - may contain errors
2 protocols using prestoblue assay
1
Aptamer Effects on Cancer Cell Viability
2
Cell Viability Assays of Encapsulated Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Live/dead assay was performed using ethidium homodimer-1 and calcein-AM (Invitrogen) according to manufacturer's protocol at 2-4 h post-encapsulation, microphotographed (Nikon E600 fl uorescence microscope), and quantifi ed using ImageJ software (N = 30 microgels). Metabolic activity was assessed at day 1, 4, 7, 14, 21, and 28 using the Presto Blue assay (Invitrogen) following manufacturer's protocol. Metabolic activities were normalized to day 1. Presto Blue assay results were corroborated with MTT staining at day 1 and 7 (Sigma-Aldrich) (Figure S1, Supporting Information). MTT was added to cell-laden microgels cultures at 0.5 mg mL -1 , which were microphotographed after 2 h of incubation and quantifi ed using ImageJ software (N > 15 microgels).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!