Microglia were plated at a density of 2.5 × 104 cells/well in 48-well plates (Corning, Tewksbury, MA) and maintained in normal cell culture conditions for 24 hrs. Cells were treated with ST-compounds and [3H]thymidine (PerkinElmer, Waltham, MA) for 72 hrs at 37°C. To measure cell viability concomitantly with cell proliferation, cells were incubated with 10 μl WST-1 (Roche, Indianapolis, IN) for 60 min at 37°C prior to cell lysis. The WST-1 absorbance was measured using a SpectraCount BS10000 (PerkinElmer, Waltham, MA) at a wavelength of 450 nm. Culture media was removed, and cells were rinsed with 250 μl ice-cold PBS and lysed with 250 μl NaOH (1 M). Cell lysates were kept on ice for 10 min and then transferred to 7-ml glass scintillation vials (VWR Scientific, Brisbane, CA). Distilled H2O (250 μl) was added to each vial followed by 4 ml scintillation fluid (National Diagnostics, Atlanta GA; Ecoscint XR). Samples were vortexed and radioactivity was measured using a scintillation counter (PerkinElmer, Waltham, MA).
Wst 1
Manufactured by PerkinElmer
Sourced in United States
The WST-1 is a colorimetric assay for the quantitative determination of viable cells in proliferation, cytotoxicity, and apoptosis. It uses a tetrazolium salt that is reduced by metabolically active cells, producing a colored formazan dye that can be measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Wst 1, by Roche (5 mentions)
Enspire, by PerkinElmer (2 mentions)
48 well plate, by Corning (1 mentions)
Spectracount bs10000, by PerkinElmer (1 mentions)
Ecoscint xr, by National Diagnostics (1 mentions)
Glass scintillation vials, by Avantor (1 mentions)
Scintillation counter, by PerkinElmer (1 mentions)
3h thymidine, by PerkinElmer (1 mentions)
Prism 5, by GraphPad (1 mentions)
Victor x4, by PerkinElmer (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Cck 8, by PerkinElmer (1 mentions)
Quant it picogreen dsdna assay kit, by Thermo Fisher Scientific (1 mentions)
1420 multilabel counter, by PerkinElmer (1 mentions)
1420 multilabel counter victor3 5, by PerkinElmer (1 mentions)
Wst 1, by Merck Group (1 mentions)
Cytotox 96 non radio cytotoxicity assay, by Promega (1 mentions)
Victor 1420 multilabel plate reader, by PerkinElmer (1 mentions)
Ldh cytotoxicity detection kit, by Takara Bio (1 mentions)
Envision instrument, by PerkinElmer (1 mentions)
8 protocols using wst 1
1
Microglia Proliferation and Viability Assay
2
Cell Viability Assay for Compound 8
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rat cardiomyocyte (H9c2) (ATCC, Manassas, VA, USA) cells were cultured in DMEM supplemented with 10% fetal bovine serum, 100 U/ml of penicillin, and 100 mg/ml of streptomycin at 37 °C in a 5% CO2 humidified atmosphere. The cell culture media were changed every 2–3 days, and the cells were sub-cultured once they reached 70–80% confluence.
Cell viability was assessed using the WST1 assay. Briefly, 104 cells were seeded in 96-well plates and were treated with four different concentrations of compound8 (3, 10, 30, and 100 µM) or vehicle (DMSO 0.1%) for 24 h. At the end of each treatment, cell viability was assessed using the cell proliferation reagent WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolium]-1,3-benzene disulphonate) (Roche, Basel, Switzerland), which was cleaved to formazan in living cells. WST-1 was added at 1:10 of the total volume of wells and after 60 min of incubation at 37 °C, the absorbance was measured at 450 nm with a multiplate reader (Enspire, Perkin-Elmer, Waltham, MA, USA). Experiments were performed in triplicate and were repeated at least three times. Data analysis for concentration–response experiments was performed using GraphPad Prism 5 (GraphPad Software, La Jolla, CA, USA), and the data were normalized with respect to the vehicle values, representing 100% cell growth.
Cell viability was assessed using the WST1 assay. Briefly, 104 cells were seeded in 96-well plates and were treated with four different concentrations of compound
3
Measurement of Neuronal Metabolic Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To measure the metabolic activity of viable cells, WST-1 (Roche, Switzerland) or CCK-8 (Dojindo, Japan) assay was performed according to the manufacturer’s instructions. Primary cortical neurons were plated at a density of 3 × 103 cells/well in the 96-well plate. Next, 1 μg/mL PSM-04 or medium was added into the media for pretreatment of primary cortical neurons (DIV 7-8) 12 h before treatment with 20 μM of the oligomeric form of Aβ1-42 or F12 medium. Thereafter, 200 ng/mL BDNF was used as positive control. Then, WST-1 or CCK-8 reagent was added to each well, and primary cortical neurons were additionally incubated at 37°C in 5% CO2 for 2 h. The absorbance of control or treated samples was measured using a multi-label plate reader (PerkinElmer, VICTOR X4) at OD of 450 nm.
4
Cell Proliferation and DNA Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
(a) WST-1:
Culture medium
was gently removed, the new medium supplemented with the cell proliferation
reagent WST-1 (Roche, Switzerland) at a final concentration of 1:10
(WST-1 stock solution/total working solution) was added, and the culture
plates were incubated at 37 °C, 5% CO2 for 2 h. The
absorbance was measured at λ = 450 nm with a plate reader (PerkinElmer
1420 Multilabel Counter). All samples were measured in triplicates.
(b) Quant-iT PicoGreen dsDNA Assay Kit: The amount of dsDNA in each
sample was quantified according to the kit manual (Invitrogen, Thermo
Fisher, USA). In brief, 28.7 μL of sample, 71.3 μL of
1X PicoGreen solution, and 100 μL 1× TE were mixed and
incubated in a 96-well plate in the dark for 10 min and the fluorescence
was read with a plate reader (λexcitation = 485 nm,
λemission = 528 nm). All samples were measured in
triplicates.
Culture medium
was gently removed, the new medium supplemented with the cell proliferation
reagent WST-1 (Roche, Switzerland) at a final concentration of 1:10
(WST-1 stock solution/total working solution) was added, and the culture
plates were incubated at 37 °C, 5% CO2 for 2 h. The
absorbance was measured at λ = 450 nm with a plate reader (PerkinElmer
1420 Multilabel Counter). All samples were measured in triplicates.
(b) Quant-iT PicoGreen dsDNA Assay Kit: The amount of dsDNA in each
sample was quantified according to the kit manual (Invitrogen, Thermo
Fisher, USA). In brief, 28.7 μL of sample, 71.3 μL of
1X PicoGreen solution, and 100 μL 1× TE were mixed and
incubated in a 96-well plate in the dark for 10 min and the fluorescence
was read with a plate reader (λexcitation = 485 nm,
λemission = 528 nm). All samples were measured in
triplicates.
5
Cell Proliferation Assay with WST-1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Stable transfected cells were seeded onto 96-well microliter plates and cell proliferation was measured with the cell proliferation Reagent WST-1 (5015944001; Sigma-Aldrich) according to the manufacturer’s recommendations. The absorption of WST-1 was measured at 450 nm using a Multilabel counter 1420 Victor3 V (Perkin Elmer, Wellesley, MA, USA).
6
Cytotoxicity and Cell Proliferation Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation reagent WST-1 (Roche Diagnostics GmbH, Mannheim, Germany) was used according to the instructions from the manufacturer. LDH release was measured using the CytoTox 96-Non-Radio Cytotoxicity Assay (Promega Biotech AB, Nacka, Sweden), according to the manufacturer’s instructions. Absorbance was measured at 450 nm for WST-1 and 490 nm for LDH (VICTOR 1420 multilabel plate reader, PerkinElmer Life Sciences, Waltham, MA, USA).
7
Evaluating Compound Cytotoxicity and Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytotoxicity was evaluated using an LDH cytotoxicity detection kit (TaKaRa) according to the manufacturer’s instructions. Briefly, culture supernatant from tumor cells was harvested after 48 h culture in the presence or absence of DSF, 5-FU (Kyowa Hakko Kirin), or IKK-IV (Merck Millipore) and assayed for the concentration of lactic acid released from damaged cells. To evaluate the effects of compounds on cell growth, cells were incubated with inhibitors for 48 h, then WST-1 (Dojindo) was added to the culture for the final 30 min. An Envision instrument (PerkinElmer) was used to measure the absorbance of each well at 450 nm versus a 650-nm reference to detect the amount of formazan produced from WST-1 by mitochondrial dehydrogenases in viable cells.
8
AsPC-1 Cell Viability Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human pancreas adenocarcinoma ascites metastasis cell line AsPC-1 (Sigma-Aldrich, passage number 3, population doubling time 38-40 hr) was cultured in RPMI 1640 (Sigma-Aldrich) supplemented with 2 mM glutamine (Sigma-Aldrich), 1 mM sodium pyruvate (Sigma-Aldrich), 10% foetal bovine serum (Sigma-Aldrich), and 1% of 100 units/ml penicillin and 100 mg/ml streptomycin (Sigma-Aldrich) in tissue culture flasks at 37°C in a humidified atmosphere and 5% CO 2 .
AsPC-1 cells were seeded onto 96-well plates at a density of 10 4 cells per well. After 24 hr, to allow cell attachment, the medium was replaced in each 96-well plate, and the cells were treated for 72 hr with ERU at the concentrations of 100, 30, 10 μM, or vehicle (dimethyl sulfoxide [DMSO] 1%). At the end of each treatment, cell viability was assessed using the cell proliferation reagent WST-1 (Roche, Basel, Switzerland; Citi et al., 2018) (link), which is cleaved to formazan in living cells. WST-1 was added at a ratio of 1:10 of the total volume of the wells, and after 60 min of incubation at 37°C, the absorbance was measured at 450 nm by a multiplate reader (Enspire, Perkin-Elmer, Waltham, Massachusetts, United States).
AsPC-1 cells were seeded onto 96-well plates at a density of 10 4 cells per well. After 24 hr, to allow cell attachment, the medium was replaced in each 96-well plate, and the cells were treated for 72 hr with ERU at the concentrations of 100, 30, 10 μM, or vehicle (dimethyl sulfoxide [DMSO] 1%). At the end of each treatment, cell viability was assessed using the cell proliferation reagent WST-1 (Roche, Basel, Switzerland; Citi et al., 2018) (link), which is cleaved to formazan in living cells. WST-1 was added at a ratio of 1:10 of the total volume of the wells, and after 60 min of incubation at 37°C, the absorbance was measured at 450 nm by a multiplate reader (Enspire, Perkin-Elmer, Waltham, Massachusetts, United States).
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!