My problem with the assay is, that the positive control doesnt work properly. Briefly, 10 l of mts reagent was added directly to the wells and cell plates were incubated at 37c for a minimum of 1 hour. Cell viability assays assay guidance manual ncbi bookshelf. Deadend fluorometric tunel system promega corporation. Celltiter 96 nonradioactive cell proliferation assay.
Celltiter 96aqueous assay mts 34,5dimethylthiazol2yl53carboxymethoxyphenyl24sulfophenyl2htetrazolium, inner salt. Celltiter 96r aqueous one solution cell proliferation assay. The realtimeglo mt cell viability assaya is a nonlytic, homogeneous, bioluminescent method to measure cell viability in real time using a simple, platebased method. This convenient one solution format is an improvement over the first version of the celltiter 96 aq ueous assay, where phenazine methosulfate pms is used as the electron coupling reagent, and pms solution and mts solution are supplied. I started doing tests to determine which concentration of dmso is toxic for the cells, as i will soon go on working with phenolic extracts which i plan to disolve in dmso. Structures of mts tetrazolium and its formazan product. Promega has contributed to this body of work and has done extensive optimization of the component formulations and conditions used to create the celltiter 96 nonradioactive cell proliferation assay for eukaryotic cell proliferation, cell attachment 3,4, and apoptosis 6 studies. The conversion of mts into aqueous, soluble formazan is accomplished by dehydrogenase enzymes found. The celltiter 96 aq ueous one solution cell proliferation assay is a colorimetric method for determining the number of viable cells in proliferation, cytotoxicity or chemosensitivity assays.
Celltiter 96r aqueous one solution cell proliferation. Peroxide was measured using o xidized 2,2 azino bis3. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516 3. Sequential multiplexing of celltox green cytotoxicity assay and celltiterglo 2. Due care should be exercised with all promega products to prevent direct human contact. Since i am using 24well plate, can i remove the culture supernatant, add in fresh media with mts, and incubate. There is linearity in the dose response curve over the range of 30125 pgml rhuil15 in the assay.
The celltiter 96 aq ueous assay is composed of solutions of a novel tetrazolium compound 34,5dimethylthiazol2yl53. The celltiter 96 aqueous one solution assay promega is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The conversion of mts into aqueous, soluble formazan is accomplished by dehydrogenase enzymes found in metabolically active cells. The mts tetrazolium is similar to the widely used mtt tetrazolium, with the advantage that the formazan product of mts reduction is soluble in cell culture medium. Email promega technical services if you have questions on use of this system. It can be purchased separately and is provided in powdered form. Promega celltiter 96 aqueous one solution cell proliferation. Celltiter 96 aqueous nonradioactive cell proliferation assay. Mts dye based colorimetric ctll2 cell proliferation assay. Introduction the turner biosystems veritas microplate luminometer in combination with promega s kinaseglo luminescent kinase assay kit provides a convenient, rapid, and sensitive procedure for quantifying the amount of atp remaining in solution after a kinase reaction. Pes has enhanced chemical stability, which allows it to be combined with mts to form a stable solution. In addition to being more convenient, rapid assays reduce the chance of artifacts caused by interaction of the test compound with assay chemistry. Promega corporation is a leader in providing innovative solutions and technical support to the life sciences industry.
The celltiterglo assay figure 2 promega corporation, madison, wi is an example of the latest bioluminescence assays, which measures cellular atp content using the enzyme luciferase hannah. Hello,i am doing mts assays working with hl60 cells. The mts assay reagent is composed of mts and the electron coupling agent phenazine methosulfate. Sv total rna isolation system quick protocol card, 9fb025.
Absorbance was measured at 490 nm on a microplate reader. Celltiter 96 aqueous one solution cell proliferation assay mts. Feb 24, 2010 mts assay posted in tissue and cell culture. A colorimetric method for determining the number of viable cells. Mmlv reverse transcriptase in 1x reaction buffer for one hour at 37c. Mts cell viability assay of upcyte hepatocyte cultures. The nonlytic nature of this assay allows the cells to be used for further downstream applications, including multiplexing with a variety of assay chemistries. Our teams are in regular contact with suppliers and distributors worldwide and are taking all steps necessary to address both demands for diagnostic tools and reliable delivery of all products as quickly as possible. Promega products contain chemicals which may be harmful if misused. One solution cell proliferation assay system technical bulletinpdf 737 kb english. The assay of this genetic reporter is rapid, sensitive and quantitative. Choosing the right cellbased assay for your research. Following incubation, the supercoiled dna is visualized on an ethidium bromidestained agarose gel to verify the absence of visible nicking or cutting analysis on 0.
Product components and storage conditions product size cat. Measurements were made according to manufacturers instructions. A simple highcontent cell cycle assay reveals frequent. Sv total rna isolation system quick protocol card, 9fb025 author. Our product portfolio of over 4,000 products supports a range of life science work in cell biology, dna, rna and protein analysis, drug development, human identification and molecular diagnostics. Aug 31, 2009 the mts dye based colorimetric assay using aqueous one 96 cell titer assay kit from promega corporation is qualified for quantitative estimation of il15 dependent cell proliferation activity. The celltiter 96 aq ueous one solution reagent contains a tetrazolium compound 34,5dimethylthiazol2yl53carboxymethoxyphenyl24sulfophenyl2htetrazolium, inner salt. The pgl2 vectors carry the coding region for firefly photinus pyralis luciferase, used to monitor transcriptional activity in transfected eukaryotic cells.
Promega products are warranted to meet or exceed the stated specifications. Animals were handled daily in a quiet room for up to 7 days before the treatments. Promega produces a complete portfolio of homogeneous assays assays that can be performed in your cell culture plates that are designed to meet a variety of experimental. Mmlv reverse transcriptase certificate of analysis 9pim170.
The data were presented as number of cells, the percentage of viable cells %, inhibition proliferation of. These assays are used for measuring the results of cell proliferation, testing. Assays are performed by adding a small amount of the celltiter 96 aqueous one solution reagent directly to culture wells, incubating for 14 hours and then recording absorbance at 490nm with a 96well plate reader. Mts reduction resazurin reduction a tp a tp p arameter measured 800 cells200 cells 390 cells50 cells 40 cellsn. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. Celltiter 96 aqueous nonradioactive cell proliferation. Promega celltiter 96 aq ueous nonradioactive cell proliferation assay kit mts homogeneous, colorimetric cell viability assay.
Celltiter 96 aq ueous nonradioactive cell proliferation assay. The one solution contains mts compound and an electron coupling reagent pes. Mts cell viability assay the celltiter 96 aq ueous assays are based on the reduction of the tetrazolium salt, mts, to a colored formazan compound by viable cells in culture. Lipgnrtd were mixed with lipgnrr6g with a volume ratio of 9. Each promega product is shipped with documentation stating specifications and other technical information. For more detailed information, refer to promega technical bulletin. Just as the manual states, i have found that lymphocyte cell lines leukemic cell lines require a lot more cells than other. Pureyield plasmid midiprep system protocol promega. These factors may be cisacting, such as promoters and enhancers, or transacting, such as various dnabinding factors. Cells were treated with or without compound saha or. Shop online for a wide selection of promega celltiter 96 aqueous one solution cell proliferation assay mts homogeneous, colorimetric cell viability assay.
Instructions for use of products a2492, a2495, a2496. Dualluciferase reporter assay system protocol promega. Mts is bioreduced by cells into a formazan product that is soluble in tissue culture medium. Promega corporation2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 6082744330 fax 6082772516. Cell viability protocols and applications guide promega. Celltiter 96 aqueous nonradioactive cell proliferation assay technical bulletinpdf 330 kb english. The plate was incubated in 5% co 2 at 37c for 4 hours. Celltiter 96 aqueous one solution cell proliferation assay. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 6082744330 fax 6082772516. The absorbance of the formazan at 490nm can be measured directly from 96well assay plates without additional processing.
Certificate of analysis promega corporation 2800 woods hollow road madison, wi 537115399 usa telephone 6082744330 toll free 8003569526 fax 6082772516 internet. Absorbance was measured at 490 nm on a spectramax plus384 reader molecular devices. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516 5. Cell viability was assessed with an mts assay kit promega corporation, madison, wi. The pureyield plasmid midiprep system is designed to isolate highquality plasmid dna for use in eukaryotic transfection and in vitro expression experiments. Celltiter 96 aq ueous mts reagent powder is a novel tetrazolium compound for use in colorimetric assays to determine the number of viable cells in proliferation, cytotoxicity or chemosensitivity assays.
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