In order determine end-point cell viability, cell titre blue (Promega, Southampton, UK) was added to each well following manufacturer’s instructions and incubated for 1.5 h, before the fluorescence of each well was measured using a Clariostar plate reader. All stats were performed on the raw data but were presented by normalising to scRNA controls to give relative viability.
Celltitre blue
Manufactured by Promega
Sourced in United Kingdom, United States
CellTiter-Blue is a cell viability assay kit that measures the metabolic activity of cells. It uses a resazurin-based solution that is converted to a fluorescent product by metabolically active cells. The amount of fluorescence generated is proportional to the number of viable cells present in the sample.
Automatically generated - may contain errors
Lab products found in correlation
Modulus 2 9300 062, by Promega (1 mentions)
Clariostar plus, by BMG Labtech (1 mentions)
L ascorbic acid 2 phosphate, by Fujifilm (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Fluostar omega plate reader, by BMG Labtech (1 mentions)
Beta apo 8 carotenal, by Merck Group (1 mentions)
Mitosox red, by Thermo Fisher Scientific (1 mentions)
Carbonyl cyanide p trifluoromethoxyphenylhydrazone fccp, by Merck Group (1 mentions)
Antimycin a, by Merck Group (1 mentions)
1 palmitoyl 2 5 oxo valeroyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
Silica gel 60, by Merck Group (1 mentions)
Cell proliferation kit 2 xtt, by Roche (1 mentions)
Dna prep coulter reagent kit, by Beckman Coulter (1 mentions)
Annexin 5 fitc pi kit, by Beckman Coulter (1 mentions)
12 protocols using celltitre blue
1
Cell Viability Assay Using CellTiter-Blue
2
Assessing bLF Cytotoxicity in A549 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549 cells were seeded in 75 µL complete medium per well in a black, flat bottomed, 96-well tissue-culture-treated plate (Fisher Scientific, Loughborough, UK) and incubated under tissue culture conditions for 24 h. Cells were then treated by mixing in an additional 25 µL of drug/diluent control in complete medium. bLF was added to the cells at a maximal final concentration of 4 mg/mL and as a positive toxic control, staurosporine was alternatively added at a maximum final concentration of 50 µg/mL, and both were serially diluted to 1:1 in complete medium; 0.2% Triton X-100 was used as an additional positive control for cell death. Cells were incubated for 20 h under tissue culture conditions before 20 µL CellTitre Blue (Promega, Southampton, UK) was added and returned to tissue culture conditions for 4 h. Fluorescence intensity was measured using a plate reader (Tecan, Theale, UK). A one-way ANOVA was performed to determine significant changes in viability.
3
Cell Viability Determination Assay
4
CellTitre-Blue Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A CellTitre-Blue® assay (Promega, UK) was used to evaluate cell viability. Scaffolds were placed in a new 48 well suspension plate and washed 3 times in PBS, 480 µl of stock solution (5:1, media/assay) was added to each scaffold and incubated for 2 h. Fluorescence was read using a microplate reader (Modulus II 9300-062, Turner Biosystems) at Ex 520 nm Em 580–640 nm, N = 4 independent replicates. A standard curve was used to estimate cell number.
5
Ovarian Cancer Cell Lines Viability Assays
6
MNV-1 Culture Assays and Cytotoxicity Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MNV-1 (CW1) culture assays were performed as previously described14 (link). Replication was examined using plaque assays58 ,64 (link),65 (link). Cytotoxicity was examined using a CellTitre Blue (Promega, Madison, WI, USA) viability assay as described previously14 (link).
7
Cell Viability Quantification on Scaffolds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability was determined using CellTitre-Blue® assay (Promega), conducted as per the manufacturer’s instructions at 24 h, Day 3 and Day 7. In short, cell seeded scaffolds (N = 5) were transferred to a fresh suspension plate with 400 µL fresh media and 100 µL CellTitre-Blue and incubated at 37°C, 5% CO2 for a total of 3 h. 100 μL of each sample was transferred to a 96 well plate and fluorescence measured at 560/590 nm using a Clariostar® Plus microplate reader (BMG LABTECH).
8
Osteoblastic Differentiation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hBSMC‐TERT and primary hMSC were plated at 20,000 cells per cm2 in osteoblastic induction medium (OIM) as previously reported.21 In brief, MEM was supplemented with 10 mM β‐glycerolphosphate (Calbiochem, EMD Millipore, Billerica, MA, USA), 50 µg/ml L‐ascorbic acid‐2‐phosphate (Wako Chemicals, Richmond, VA, USA), 10 nM dexamethasone (Sigma‐Aldrich, St. Louis, MO, USA), and 10 nM calcitrol (LEO Pharma, Madison, NJ, USA) for a maximum of 15 days. Medium was replaced every 3 days.
Alkaline phosphatase (ALP) activity was measured at day 6 of OIM using Cell Titre Blue (Promega, Madison, WI, USA) to measure cell viability and p‐nitro phenyl phosphate (pNP) to measure ALP activity as previously described.21 Absorbance was measured on a FLUOstar Omega plate reader (BMG LabTech, Cary, NC, USA) and ALP activity corrected for cell viability (n = 6).
Alkaline phosphatase (ALP) activity was measured at day 6 of OIM using Cell Titre Blue (Promega, Madison, WI, USA) to measure cell viability and p‐nitro phenyl phosphate (pNP) to measure ALP activity as previously described.
9
Neuroblastoma SH-SY5Y Cell Carotenoid Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human neuroblastoma SH-SY5Y cells were purchased from American Type Culture Collections, Manassas, USA. Authentic standards of lutein and beta-apo-8'-carotenal were purchased from Sigma Aldrich (Dorset, UK); zeaxanthin was purchased from Cambridge Biosciences (Cambridge, UK); carotenoid standards were kept in the dark, made up fresh and used immediately after preparation and confirmation of concentrations. 1-palmitoyl-2-(5'-oxo-valeroyl)-sn-glycero-3-phosphocholine (POVPC) was purchased from Avanti Polar Lipids (Alabaster, AL, USA); MitoSOX TM Red was purchased from Invitrogen (Fisher Scientific, Loughborough, UK); cell titre blue was purchased from Promega Corporation (Hollow Road, USA). All materials for the Extracellular Flux assays were from Agilent Technologies / Seahorse Biosciences / (Santa Clara, USA). Carbonyl cyanide p-
[trifluoromethoxy]-phenyl-hydrazone (FCCP), oligomycin, and antimycin A were from Sigma-Aldrich.
All solvents used were HPLC grade from Fisher Scientific (Loughborough, UK). All other cell culture media and chemicals were also purchased from Fisher Scientific (Loughborough, UK) unless otherwise stated.
[trifluoromethoxy]-phenyl-hydrazone (FCCP), oligomycin, and antimycin A were from Sigma-Aldrich.
All solvents used were HPLC grade from Fisher Scientific (Loughborough, UK). All other cell culture media and chemicals were also purchased from Fisher Scientific (Loughborough, UK) unless otherwise stated.
10
Assessing Cell Viability After miRNA Transfection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Assay miR transfections were examined to determine if transfection mediated >20 % loss in cell viability using Cell Titre Blue (CTB; Promega, WI) for any miR mimic or inhibitor pair. Briefly, A549 cells were transfected with miR mimic, miR inhibitor, siTOX, or mock-transfected [24 (link)]. Following 48 h incubation, the transfected cell viability was determined according to the manufacturer’s protocol. Briefly, 100 µl of media from each well was decanted and 20 µl of CTB reagent added to each well. Plates were mixed gently for 10 s and then incubated at 37 °C. 5 % CO2 for 2 h. Following incubation, the plates were gently rocked for 10 s before reading absorbance with Tecan plate reader at 570 nm with reference at 600 nm. Percent viability was calculated by comparing mock-transfected to miR-transfected (Table S3).
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!