Wst 1 cell proliferation assay kit

Manufactured by Cayman Chemical

Sourced in United States

The WST-1 cell proliferation assay kit is a colorimetric method for the non-radioactive quantification of cell proliferation and viability. The assay is based on the cleavage of the tetrazolium salt WST-1 to formazan by cellular mitochondrial dehydrogenases. The amount of formazan dye generated is directly proportional to the number of metabolically active cells in the culture.

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Lab products found in correlation

Lomustine, by Merck Group (2 mentions) Mibefradil dihydrochloride hydrate, by Merck Group (2 mentions) Multiskan go, by Thermo Fisher Scientific (2 mentions) Microplate reader, by Agilent Technologies (1 mentions) Verapamil hydrochloride v4629, by Merck Group (1 mentions) Nnc55 0396 hydrate, by Merck Group (1 mentions) Synergy ht, by Agilent Technologies (1 mentions) 96 well tissue culture microplates, by Corning (1 mentions) Nnc 55 0396 hydrate n0287, by Merck Group (1 mentions)

14 protocols using wst 1 cell proliferation assay kit

Platelet Viability Assessment using WST-1

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The water-soluble tetrazolium salt-1 (WST-1) test was used to assess platelet viability
in the two groups with the WST-1 cell proliferation assay kit (Cayman Chemical, Ann Arbor,
MI, USA) and a 96-well microplate. All PC samples were centrifuged at 1800 g for 4 minutes
and the platelets were re-suspended in phosphate-buffered saline. Platelet concentrations
of 5×10¹¹ cells/L in a 100 μL suspension were used with the addition of 10 μL
of WST-1 reagent mix, followed by incubation for 4 hours in a CO₂ incubator at
37˚C. The absorbance of the reaction was read at 450 nm as the optical density (OD) in a
microplate reader (ASYS Expert 96 UV Microplate Reader, UK). Results of the WST-1 analysis
were presented as mean ± SD.

Rajabi A., Sharifi Z., Yari F., Deyhim M, & Jalili M. (2020). The Effect of Composol Medium on miR-16 Expression during Platelet Storage up to Day 7 at Room Temperature. Cell Journal (Yakhteh), 22(4), 542-547.

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Mitochondrial Dehydrogenase Activity Assay

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WST-1 cell proliferation assay kit (WST-1, Cayman,
USA) was used to measure the activity of cellular
mitochondrial dehydrogenases in the PLTs. In this test,
the tetrazolium salt is changed to formazan by viable
PLTs; therefore, the result indicates PLTs viability rate.
Following diluted with PBS, 10×10⁶ PLTs (100 µl)
were added into each well. Accordingly, 10 µl of the
WST-1 mixture was added to each well, and the plate
was incubated at 37°C in an incubator for 4 hours. The
absorbance of the samples was measured at 450 nm in a
microplate reader (Asys Expert 96, UK).

Baghdadi V., Ranjbaran R., Yari F, & Rafiee M.H. (2022). Trehalose An Additive Solution for Platelet Concentrate to Protect Platelets from Apoptosis and Clearance during Their Storage at 4°C. Cell Journal (Yakhteh), 24(2), 69-75.

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Mitochondrial Metabolic Activity Assay for MCF-7 and Panc-1 Cells

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Mitochondrial metabolic activity of MCF-7 and Panc-1 cells after administration of GA, GA + Gn and GA + SWCNT was determined using the WST-1 Cell Proliferation Assay Kit (Cayman Chemical Company, Ann Arbor, MI, USA). This assay takes advantage of the color shift that occurs after mitochondrial dehydrogenase cleaves the tetrazolium salt, WST-1, to formazan in viable cells. Briefly, MCF-7 and Panc-1 cells were seeded on to 96-well plates at a density of 25 × 10³ cells per well and allowed to grow for 24 h before treatment with increasing concentrations of GA (0–1 μg ml⁻¹) at a constant Gn or SWCNT concentration (10 μg ml⁻¹). After 48 h incubation, 10 μl of reconstituted WST-1 mixture were added to each well and incubated for 2 h in the dark at 37 °C. The absorbance at 450 nm was measured using a microplate reader (Biotek, Winooski, VT, USA). Wells containing only medium and WST-1 reagent were measured as a background control. All conditions were run in triplicate for statistical analysis. IC₅₀ values (the concentration of the drug that inhibits cell viability by 50%) were calculated based on the results of the WST-1 assay.

Saeed L.M., Mahmood M., Pyrek S.J., Fahmi T., Xu Y., Mustafa T., Nima Z.A., Bratton S.M., Casciano D., Dervishi E., Radominska-Pandya A, & Biris A.S. (2014). Single-walled carbon nanotube and graphene nanodelivery of gambogic acid increases its cytotoxicity in breast and pancreatic cancer cells. Journal of applied toxicology : JAT, 34(11), 1188-1199.

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Evaluating Lactobacilli EPSs' Impact on HT-29 Cell Proliferation

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Impact of EPSs from lactobacilli on HT-29 cell proliferation was evaluated using a WST-1 cell proliferation assay kit (Cayman Chemical Company, Ann Arbor, Michigan, USA). The lyophilized EPSs were dissolved in distilled water and filtered using a 0.2 µm syringe filter prior to analyses. HT-29 cells were seeded into a 96-well plate at a density of 1 × 10⁴ cells/well and treated with EPSs at a final concentration of 400 µg/ml followed by 24 h or 48 h incubation at 37 °C with 95% air and 5% CO₂. After incubation, 10 μl of the WST-1 mixture was added to each well and the plates were incubated for 2 h at 37 °C with 95% air and 5% CO₂. Formation of formazan was measured at 450 nm by a microplate reader (Epoch, Biotek, Winooski, VT, USA) and the absorbance was correlated with the cell number. The anti-proliferative effect was evaluated by comparing to viability of the treated samples with the untreated control (ultrapure water and DMEM mix without test sample for EPS). The percentage of viability was calculated as follows:

% Viability = (Absorbance Sample / Absorbance Control) \times 100

Tukenmez U., Aktas B., Aslim B, & Yavuz S. (2019). The relationship between the structural characteristics of lactobacilli-EPS and its ability to induce apoptosis in colon cancer cells in vitro. Scientific Reports, 9, 8268.

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Cell Viability Assessment using WST-1 Assay

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Cell viability was assessed with a WST-1 Cell Proliferation Assay kit (Cayman Chemical), which measures the enzymatic conversion of WST-1 to formazan by cell mitochondrial dehydrogenases present in viable cells. A549 cells were seeded in a 96-well format at a density of 5000 cells/well. Cells were then transfected with 100 or 50 nM of different miRNAs using INTERFERin as described above. Cell viability was measured 48 h post-transfection as per the manufacturer's instructions. Briefly, cells were incubated with 10 μL of WST-1 for 30 min—1 h at 37°C, 5% CO₂. The absorbance was read at 450 nm using a microplate reader. Optical density (OD) is shown as a percent of viable control cells, No miR. Assays were performed in independent triplicates.

Cornett A.L, & Lutz C.S. (2014). Regulation of COX-2 expression by miR-146a in lung cancer cells. RNA, 20(9), 1419-1430.

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PSC Viability Assay by WST-1

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The viability of PSCs treated by DMSO or SFN (5, 10, 15, or 20 μM) in 96-well plates was determined using a WST-1 cell proliferation assay kit (Cayman Chemical, Hamburg, Germany). For each well, 10 µL WST-1 reagent was added and cells were cultured at 37 °C for two hours. The absorbance at 450 nm was assessed and all readouts were normalized with the background value.

Zhang R., Neuhoff C., Yang Q., Cinar M.U., Uddin M.J., Tholen E., Schellander K, & Tesfaye D. (2022). Sulforaphane Enhanced Proliferation of Porcine Satellite Cells via Epigenetic Augmentation of SMAD7. Animals : an Open Access Journal from MDPI, 12(11), 1365.

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HUVEC Proliferation on Scaffold Mats

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HUVECs were cultured
in EBM-2 supplemented with growth factor kit and maintained at 37
°C and 5% CO₂. Prior to cell seeding, the scaffolds
were cut into 0.5 cm × 0.5 cm × 0.2 cm and sterilized in
70% ethanol for 10 min, followed by 10 min of UV exposure. HUVEC cells
were seeded at 150 000 cells per mat. The cells were allowed
to attach for 1 h; then, 200 μL of fresh medium was added
to each well. The WST-1 cell proliferation assay kit was used according
to the manufacturer’s instructions to quantitatively evaluate
proliferation on seeded mats (n = 5), (Cayman, MI).
Cells seeded on tissue culture plates (TCPs) served as the control.
At predetermined time points (1, 3, 5, and 10 days), the medium was
removed, samples washed with PBS twice to remove unattached cells,
100 μL of fresh medium was added, and then 10 μL of WST-1
solution was added and incubated for 2 h at 37 °C. The medium
(100 μL) was then transferred to a 96-well plate and absorbance
was measured at 450 nm. For cell attachment and cell spreading observations,
seeded mats were fixed with 70% ethanol overnight and then observed
using FE-SEM at each predetermined time point.

Hou L., Zhang X., Mikael P.E., Lin L., Dong W., Zheng Y., Simmons T.J., Zhang F, & Linhardt R.J. (2017). Biodegradable and Bioactive PCL–PGS Core–Shell Fibers for Tissue Engineering. ACS Omega, 2(10), 6321-6328.

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Cell Viability and Proliferation Assay

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Metabolic activity of cells as a marker of cell viability and proliferation was assessed by WST-1 Cell Proliferation Assay Kit (10,008,883, Cayman Chemical, MICH, USA) according to the manufacture’s recommendation. Three MB cell lines, D341, CHLA-01 and CHLA-01R, were used in this assay. Briefly, cells were seeded at a density of 30,000 cells/well in 96-well plates. Twenty-four hours later, treatment was added as shown in the “Results” section. Assay reaction was assessed 24 or 72 h after treatment. For that, 10 μL of WST-1 reagent was added to each well for 4 h, and absorption was measured using a plate reader (Multiskan Go, Thermo Fisher Scientific, Scoresby, VIC, Australia). Drugs used, including mibefradil dihydrochloride hydrate (M5441), NNC55-0396 hydrate (N0287), verapamil hydrochloride (V4629), nifedipine (N7634), vincristine (V0400000) and lomustine (L5918), were purchased from Sigma-Aldrich.

Sedeeq M., Maklad A., Dutta T., Feng Z., Wilson R., Gueven N, & Azimi I. (2022). T-Type Calcium Channel Inhibitors Induce Apoptosis in Medulloblastoma Cells Associated with Altered Metabolic Activity. Molecular Neurobiology, 59(5), 2932-2945.

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Mitochondrial Activity Assay for Platelets

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To measure the mitochondrial activity of PLTs, we used
the WST-1 cell proliferation assay kit (WST-1, Cayman,
USA). In this method, tetrazolium salt is changed to
formazan in viable cells by cellular mitochondrial
dehydrogenases. Here, PLTs were diluted with phosphate
buffered saline (PBS) and 10×10⁶ PLTs (100 μl) were
added into each well. Then, 10 μl of WST-1 blend solution
was added to each well and the plate was incubated for 4
hours at 37˚C in a CO₂ incubator. The absorbance of the
wells was measured using a microplate reader at 450 nm.

Baghdadi V., Yari F., Nikougoftar M, & Rafiee M.H. (2019). Platelets Apoptosis and Clearance in The Presence of Sodium Octanoate during Storage of Platelet Concentrate at 4˚C. Cell Journal (Yakhteh), 22(2), 212-217.

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Cytotoxicity of Extracts on Cell Lines

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Human breast epithelial cells (MCF-12A, ATCC CRL-10782) (Vorster et al, 2012 (link)) human breast cancer cells (SKBR 3, ATCC HTB-30) (Quirantes-Piné et al, 2013 ) and human cervix cancer cells (HeLa, ATCC CCL-2) (Lu et al, 2010 (link)) were used for cytotoxicity tests and the cytotoxicity of 500 and 1000 µg.mL⁻¹ of extracts were determined according to the protocol provided by WST-1 Cell proliferation assay kit (Cayman).

Onbasli D, & Yuvali G. (2020). In vitro medicinal potentials of Bryum capillare, a moss sample, from Turkey. Saudi Journal of Biological Sciences, 28(1), 478-483.

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