Wst 1 kit

Manufactured by Roche

Sourced in Germany, Switzerland, United States

The WST-1 kit is a cell proliferation and viability assay. It is a colorimetric assay for the quantitative determination of cell proliferation and viability in cell populations. The assay is based on the cleavage of the tetrazolium salt WST-1 to a colored formazan dye by mitochondrial dehydrogenases, which directly correlates with the number of metabolically active cells in the culture.

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Lab products found in correlation

Spectramax m5 spectrophotometer, by Molecular Devices (2 mentions) Fluostar omega plate reader, by BMG Labtech (2 mentions) Elisa reader, by Tecan (1 mentions) Caspase 3 cpp32 colorimetric assay kit, by Abcam (1 mentions) Glyceraldehyde 3 phosphate dehydrogenase gapdh antibody, by Santa Cruz Biotechnology (1 mentions) Nanoquant spectrophotometer, by Tecan (1 mentions) Curcumin, by Merck Group (1 mentions) Wst 1, by Roche (1 mentions) Ldh kit, by ScienCell (1 mentions) 96 well plate, by SPL Life Sciences (1 mentions) El808 ultra microplate reader, by Agilent Technologies (1 mentions) Epoch microplate reader, by Agilent Technologies (1 mentions) Trypan blue, by Merck Group (1 mentions) 96 well plate reader, by Tecan (1 mentions) Gemini em, by Molecular Devices (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Heregulin, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Ag1478, by Bio-Techne (1 mentions) Cell culture media, by Thermo Fisher Scientific (1 mentions)

44 protocols using wst 1 kit

Cell Proliferation Analysis of MG-63 Cells

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MG-63 was tested using the cell proliferation reagent WST-1-Kit (Roche Diagnostics GmbH, Mannheim, Germany). The conversion of tetrazolium salt via mitochondrial succinate-dehydrogenase-vital cells to colored formazan was measured spectrophotometrically. For each day three scaffolds were examined. The Assay was repeated three times.
The medium was removed from the microtiter plates after 3, 7, 14, and 21 days and the sample bodies were transferred to a new 24-well-plate. Both the original and the new well plate were then supplemented with 600 µL phenol-free standard medium (Gibco, Paisley, UK) and 60 µL WST-reagent. The plates were then incubated for 2 h at 37 °C and 5% CO₂. Finally, 4 × 100 µL of the supernatant was pipetted out of every well into a 96 well plate. The absorption of the liquid at a wavelength of λ = 450 nm and λ = 600 nm reference was measured via an ELISA-Reader (Tecan Deutschland GmbH, Crailsheim, Germany).

Burtscher S., Krieg P., Killinger A., Al-Ahmad A., Seidenstücker M., Latorre S.H, & Bernstein A. (2019). Thin Degradable Coatings for Optimization of Osteointegration Associated with Simultaneous Infection Prophylaxis. Materials, 12(21), 3495.

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Apoptosis Signaling Pathway Assay

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A WST‐1 kit was purchased from Roche Applied Sciences. A caspase‐3/CPP32 colorimetric assay kit was purchased from BioVision. The antiphospho‐JNK and JNK antibodies were purchased from Cell Signaling Technology. The antiphospho‐p38, p38 MAPK, and glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) antibodies were purchased from Santa Cruz Biotechnology. Sigma‐Aldrich was the manufacturer of all other chemicals.

Yeh Y., Liang C., Chen M., Tsai F., Lin Y., Lee M., Wu J, & Kuo C. (2019). Apoptotic effects of hsian‐tsao (Mesona procumbens Hemsley) on hepatic stellate cells mediated by reactive oxygen species and ERK, JNK, and caspase‐3 pathways. Food Science & Nutrition, 7(5), 1891-1898.

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ERβ2 Modulates Cell Proliferation

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3000 cells per well were seeded in 96-well plates 24 h after ERβ2 or control plasmid transfection and 48 h after ERβ2 siRNA or control siRNA transfection. Cell proliferation was determined after 1, 2, 3 or 4 days using the WST-1 kit (Roche, Basel, Switzerland) according to the instructions of the manufacturer. The absorbance was measured at 450 nm using NanoQuant spectrophotometer (Tecan, Männedorf, Switzerland).

Bialesova L., Xu L., Gustafsson J.Å., Haldosen L.A., Zhao C, & Dahlman-Wright K. (2017). Estrogen receptor β2 induces proliferation and invasiveness of triple negative breast cancer cells: association with regulation of PHD3 and HIF-1α. Oncotarget, 8(44), 76622-76633.

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Cell Viability Assays for Cancer Cell Lines

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Cell survival assays were performed on human alveolar adenocarcinoma A549, murine melanoma B16-F10, human colon adenocarcinoma Caco-2, human colorectal adenocarcinoma HT-29 and triple negative human breast adenocarcinoma MDA-MB-231 cell lines treated with complexes 1 and 2, cisplatin, bipy, and BC, as previously described.^{13 (link)} Briefly, DMEM containing 10% FBS and 1.5% pen-strep at 37 °C with 5% CO₂ was used to maintain cells. The latter were seeded and allowed to adhere at a concentration of 10⁴ cells per well in a 96 well-plate. Drugs were added to the media at different concentrations using 3-fold serial dilution. Cells were incubated with drugs for 12 h followed by 35 min exposure to light (LED Engin, 460 nm peak wavelength and 100 mW cm⁻² power output) or else the plates were left in the dark. Cell survival was measured using WST-1 kit (Roche©) after 72 h incubation. Three independent experiments were performed, each comprised of triplicate measurements. To test the activity of complex 1 on normal cells, the isolated MSCs were treated with the complex using the same conditions described for cancer cells.

Mehanna S., Mansour N., Audi H., Bodman-Smith K., Mroueh M.A., Taleb R.I., Daher C.F, & Khnayzer R.S. (2019). Enhanced cellular uptake and photochemotherapeutic potential of a lipophilic strained Ru(ii) polypyridyl complex. RSC Advances, 9(30), 17254-17265.

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Evaluating Cell Proliferation with CMO and CT

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Tumor cells: M-406 (5×10⁴ cells) was cultured with different CMO (100 μl) and with RPMI (Control group, reference value, 100%).
Mononuclear cells: MO (5×10⁴ cells) from naïve CBi (MO-CBi N), CBi⁻ (MO-CBi⁻ N) and CBi/L (MO-CBi/L N) mice were cultured with CT (100 μl).
Cells were incubated for 24 h, at 37°C and 5% of CO₂ and cell proliferation was evaluated with WST-1 kit (Roche, Argentina) (Figure 2).

Pagura L., Cáceres J.M., Cardinale A., Scharovsky O.G., Masso R.J., Zacarías-Fluck M.F., Rico M.J, & Rozados V.R. (2014). A mammary adenocarcinoma murine model suitable for the study of cancer immunoediting. Journal of Biomedical Science, 21(1), 52.

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Nanoparticle Viability Assay in Dendritic Cells

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For viability studies, we used a WST-1 kit (Roche). DCs were seeded in 96-well plates at 10,000 cells/well and allowed to adhere overnight. Once attached, the DCs were treated with 1, 10, 100, and 200 μg/ml of AuNR/Ag or PNVs in a final volume of 100 µL and incubated for an additional 24 hours at 5% CO₂, 37 °C, 100% humidity. After incubation, 10 µl of the WST-1 reagent was added to each well, and after 2 hours, the absorbance was read at 420–480 nm, similarly to how we have described before^{37 (link), 38 (link)}.

Vang K.B., Safina I., Darrigues E., Nedosekin D., Nima Z.A., Majeed W., Watanabe F., Kannarpady G., Kore R.A., Casciano D., Zharov V.P., Griffin R.J., Dings R.P, & Biris A.S. (2017). Modifying Dendritic Cell Activation with Plasmonic Nano Vectors. Scientific Reports, 7, 5513.

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Doxorubicin Cytotoxicity Assay

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Cells in the logarithmic growth period were cultured in a 96‐well plate at a density of 2 × 10⁵ cell/mL in a 0.1 mL volume for 6 h. Doxorubicin of variable concentrations (0.1, 0.2, 0.3, 0.4 and 0.5 μg/mL) was added to the experiment group, while the control group was cultured with the same volume of culture medium in a 5% CO₂ incubator at 37℃ for 24 h. A minimum of 3 parallel wells were set in each group. Cell viability was detected using a WST‐1 kit (Roche Applied Science, Upper Bavaria, Germany). One hour before each measurement, 100 µL Dulbecco’s modified Eagle’s medium and 10 µL WST‐1 solution were added successively, the absorbance (A) value at the wavelength of 450 nm was documented using a microplate reader. The inhibitory concentration IC₅₀ was calculated to compare the resistance.¹⁷ (link)

Li L., Gao Z., Zhao L., Ren P, & Shen H. (2021). Long non‐coding RNA LINC00607 silencing exerts antioncogenic effects on thyroid cancer through the CASP9 Promoter methylation. Journal of Cellular and Molecular Medicine, 25(16), 7608-7620.

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Cell Viability Assessment of TMJ Disc Cells

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Cell viability of the preconditioned experimental groups was measured after 48 hours using the WST-1 kit (Roche Molecular Biochemicals, Mannheim, Germany). Water-soluble tetrazolium salt, 4-(3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio)-1, 3-benzen disulfonate (WST-1), has been demonstrated to be a simple and rapid measurement of cell viability with extremely low cytotoxicity. A ten percent working solution was made by mixing one part volume of the cell viability reagent WST-1 with nine parts volume of media. Quantification of the formazan dye produced by metabolically active cells was done via a scanning multi-well spectrophotometer (420-480nm) ^{23 (link)}. Absorbance values collected from cells cultured at 25 mM glucose and 21% oxygen level were considered the control measurement due to the initial in vitro expansion culture conditions. All other absorbance values from other cell culture conditions were normalized to the control. A standard curve was performed to show the relationship between different numbers of seeded porcine TMJ disc cells and absorbance.

Cisewski S.E., Zhang L., Kuo J., Wright G.J., Wu Y., Kern M.J, & Yao H. (2015). The Effects of Oxygen Level and Glucose Concentration on the Metabolism of Porcine TMJ Disc Cells. Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society, 23(10), 1790-1796.

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Evaluating Periosteal Cell Viability

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Periosteal cells were cultured inside conventional 24-well plates and awaited to adhere overnight. The cells were exposed to the same antibiotics concentrations used for the tests inside the xCELLigence system on the next day. The culture medium was then refreshed every third day until the day 7 after drug exposure when cell growth was analyzed using a WST-1 kit (Roche, Basel, Switzerland). By using the colorimetric WST-1 assay that was based on the cleavage of tetrazolium WST-1 into orange formazan by mitochondrial dehydrogenases in viable cells, the level of orange formazan increases when mitochondrial activity increases and can be quantified through an E LISA Reader (MWG-Biotech, Ebersberg, Germany) at 450 nm with a reference wavelength of 245 nm.

Chiu C.H., Lei K.F., Chan Y.S., Ueng S.W, & Chen A.C. (2019). Real-time detection of antibiotics cytotoxicity in rabbit periosteal cells using microfluidic devices with comparison to conventional culture assays. BMC Musculoskeletal Disorders, 20, 339.

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Curcumin Inhibits hRPE Cell Proliferation

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The WST-1 kit (Roche, Indianapolis, IN, USA) was used to detect RPE cell proliferation and viability following the manufacturer’s instructions. Briefly, the hRPE cells at passage 5 who had achieved a satisfactory growth were used to prepare a single-cell suspension. The cells were seeded onto 96-well plates (5,000 cells/well in 100 μl) and incubated at 37°C with 5% CO₂. The cells were treated with 5, 10, 15, 20 μg/ml curcumin (dissolved in DMSO; Sigma-Aldrich, St. Louis, MO, USA) or DMSO. After 24, 48 and 72 h, 10 μl of WST-1 solution were added to each well, followed by incubation for 1–4 h. The optical density (OD) was measured at 450 nm to determine the optimal concentration of the curcumin-mediated inhibition of proliferation.

SUN Y, & YOU Z.P. (2014). Curcumin inhibits human retinal pigment epithelial cell proliferation. International Journal of Molecular Medicine, 34(4), 1013-1019.

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