Xtt solution

Manufactured by Sartorius

Sourced in Israel

The XTT solution is a colorimetric assay reagent used for the quantitative determination of cell viability and proliferation in cell culture studies. It contains the tetrazolium salt XTT, which is reduced by metabolically active cells to form a colored formazan product that can be detected spectrophotometrically.

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Lab products found in correlation

Model 3550, by Bio-Rad (2 mentions) Prism 8, by GraphPad (2 mentions) Genios plate reader, by Tecan (1 mentions)

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XTT reaction solution (3 citations)

7 protocols using xtt solution

Cytotoxicity Assay for MDA-MB-231 Cells

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MDA-MB-231 cells were re-suspended to a concentration of 3 × 10⁴ cells/ml in DMEM/10% FBS and plated at 100 μl/well in triplicates in flat bottom 96 well microtiter plates. Following overnight incubation, inhibitors or DMSO as control were added to a final concentration of 10 μM and cells were followed every 24 hours for 72 hours total. Activated XTT solution was prepared according to the manufacturer’s instructions (Biological Industries) and added to cells. Cells were then incubated at 37° C for 4 hours. Specific absorbance was calculated using the formula:
Specific absorbance = A₄₇₀ nm (Test) – A₄₇₀nm (Blank) - A₆₆₀ nm (Test).

Meirson T., Genna A., Lukic N., Makhnii T., Alter J., Sharma V.P., Wang Y., Samson A.O., Condeelis J.S, & Gil-Henn H. (2018). Targeting invadopodia-mediated breast cancer metastasis by using ABL kinase inhibitors. Oncotarget, 9(31), 22158-22183.

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Cell Proliferation Assay of HUCMSCs

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An XTT kit was used for the cell proliferation assay. Different FBS-cultured HUCMSCs were seeded at a density of 2 × 10³ cells/well in a 96-well plate. The final volume was 100 μL of culture medium in one well. To proceed with the proliferation assay, the cells were incubated with 150 μL XTT solution (Biological Industries, Kibbutz Beit Haemek, Israel) at 37 °C for 3 h. A microplate reader (Model 3550, Bio-Rad, Hercules, CA, USA) was used for the absorbance reading (450 nm). Proliferation curves were expressed as optical density values and were constructed on days 0, 3, and 7.

Chang Y.H., Wu K.C, & Ding D.C. (2022). Chondrogenic Potential of Human Umbilical Cord Mesenchymal Stem Cells Cultured with Exosome-Depleted Fetal Bovine Serum in an Osteoarthritis Mouse Model. Biomedicines, 10(11), 2773.

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Chondrocyte Proliferation Quantification

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Chondrocytes were plated in a 96-well microtiter plate at a density of 2 × 10³ cells per well in a final volume of 100 μl of DMEM/F12 (1:1). After treatment with the above four conditions, the cells were incubated with XTT solution (Biological Industries), 150 μL for 3 h at 37°C in accordance with the manufacturer's instructions. The absorbance was read at 450 nm in a microplate reader (Bio-Rad Model 3550, Hercules, CA, USA). Growth curves expressed by the optical density values were constructed.

Chang Y.H., Wu K.C., Liu H.W., Chu T.Y, & Ding D.C. (2018). Human umbilical cord-derived mesenchymal stem cells reduce monosodium iodoacetate-induced apoptosis in cartilage. Tzu-Chi Medical Journal, 30(2), 71-80.

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XTT-based Cell Proliferation Assay

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Cell proliferation was assessed using 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) assay. The tested cells were plated in one well of a 96-well at a density of 2 × 10³ cells with 100 μL culture medium. The optical density of cells was measured on days 0, 3, 5, and 7. The tested cells were incubated with 150 μL XTT solution (Biological Industries, Beit-Haemek, Israel) for 3 h at 37 °C in an incubator. A microplate reader (Bio-Rad Model 3550, Hercules, CA, USA) was used to measure the optical density at 450 nm. The optical density values at each time point were used to construct proliferation curves of the tested cell lines.

Chang Y.H., Wu K.C., Wang K.H, & Ding D.C. (2023). Effects of the Overexpression of Progesterone Receptors on a Precancer p53 and Rb-Defective Human Fallopian Tube Epithelial Cell Line. International Journal of Molecular Sciences, 24(14), 11823.

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Biofilm and Planktonic Cell Viability Assay

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Prior to each assay, XTT solution (Biological Industries) was thawed and mixed with a N-methyl dibenzopyrazine methyl sulfate (PMS) (Biological Industries) solution at 50∶1 (v/v). The biofilms were washed and then incubated with 60 µl of the XTT–PMS solution in 2 ml of PBS. To determine planktonic cell viability, 60 µl of the XTT–PMS solution was added to the supernatants aspirated from the biofilms and placed in a new 96-well plate. Plates containing biofilms as well as supernatants were then incubated in the dark for 3 h at 37°C. Following incubation, the color change in the solution was measured spectrophotometrically at 492 nm using a GENios plate reader (Tecan, Salzburg, Austria).

Feldman M., Al-Quntar A., Polacheck I., Friedman M, & Steinberg D. (2014). Therapeutic Potential of Thiazolidinedione-8 as an Antibiofilm Agent against Candida albicans. PLoS ONE, 9(5), e93225.

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Tomentosin's Cytotoxic Effect on Pancreatic Cancer

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The cytotoxic effect of tomentosin on pancreatic cancer cells was evaluated by XTT analysis. PANC-1 and MIA PaCa-2 cells were seeded at 2x10 3 cell in 96-well plates. After 24 hours, varying concentrations of tomentosin (0-300 µM) were treated to cells for 24, 48 and 72 hours. At the end of the speci ed hours, XTT solution (Biological Industries, 20-300-1000) was added to wells and after approximately 4 hours, absorbance value of each well was read in a microplate reader (BioTek Epoch) at 450 nm wavelength and 630 nm reference range. % cell viability was calculated and IC 50 doses of tomentosin in PANC-1 and MIA PaCa-2 cells were determined with GraphPad Prism 8.0.2 software.

Güçlü E., Çınar Ayan İ., Dursun H.G, & Vural H. (2022). Tomentosin induces apoptosis in pancreatic cancer cells through increasing reactive oxygen species and decreasing mitochondrial membrane potential. Toxicology in vitro : an international journal published in association with BIBRA, 84.

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Tomentosin's Cytotoxic Effect on Pancreatic Cancer

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Güçlü E., Ayan İ.Ç., Dursun H.G., & Vural H.C. (2022). In vitro anticancer activity of tomentosin on human pancreatic cancer cells.

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