To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. Mitochondrial toxglotm assay technical manual tm357 promega. Hello,i am doing mts assays working with hl60 cells. Instructions for use of products g5421, g5430, g5440, g1111, g1112. Jun 06, 2018 absorbance generated in cellmediated and cell free mts assays. Product components and storage conditions product size cat. Promega celltiter 96 aq ueous nonradioactive cell proliferation assay kit mts homogeneous, colorimetric cell viability assay. The details of the preparation can be found in the manual from promega. Here we describe the usage of a tetrazolium compound which is soluble in water and cell culture medium.
Mts reagent is a tetrazolium inner salt useful for determining the number of viable cells in proliferation or chemosensitivity assays. A protocol for determining viable cell number in proliferation, cytotoxicity, cell attachment, chemotaxis and apoptosis assays. Promega corporation 2800 woods hollow road madison, wi 537115399 usa. My problem with the assay is, that the positive control doesnt work properly. Pms solution with the mts solution according to the manufacturers instructions promega, g5421. Inhibition of atpase activity by the celltiterglo reagent. Antioxidantrich extracts of terminalia ferdinandiana. The realtimeglo mt cell viability assay is a nonlytic, homogeneous, bioluminescent method to measure cell viability in real time using a simple, platebased method. Page 4 promega corporation2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 6082744330 fax 6082772516. Measurements were made according to manufacturers instructions. Promega corporation 2800 woods hollow rd, madison, wisconsin 53711 rated 4. Jul 09, 2016 the details of the preparation can be found in the manual from promega. Promega s sole obligation and the customers sole remedy is limited to replacement of products free of charge in the event products fail to perform as warranted. Cell viability assay promega corporation, madison, wi, usa was used to perform the interference experiment in a cell free system.
The mts compound is bioreduced by cells into a colored formazan product that is soluble in tissue. These issues are addressed in the related sop nm interference in the mts assay. The assay of this genetic reporter is rapid, sensitive and quantitative. Each promega product is shipped with documentation stating specifications and other technical information. Limitations of mtt and mtsbased assays for measurement of. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516. The method is based on the reduction of mts tetrazolium compound by viable cells to generate a colored formazan product that is soluble in cell culture media. Promega products and technologies are used by scientists in academic and.
Celltiter 96 aqueous one solution cell proliferation assay mts. Celltiter 96 aqueous nonradioactive cell proliferation assay. B effect of serum and c effect of bovine serum albumin bsa with and without fatty acids in the cell free assay. Description continued the role of the proteasome in degrading several important regulatory proteins has led to the identification of the. Promega products are warranted to meet or exceed the stated specifications. The gel shift assay core system includes sufficient hela nuclear extract to perform 20 control reactions. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 6082744330 fax 6082772516. Mitochondrial toxglotm assay technical manual tm357. The mts tetrazolium compound owens reagent is bioreduced by cells into a colored. The resul ting mixture can be aliquoted and stored at 20 c before use. Mts is bioreduced by cells into a formazan product that is soluble in tissue culture medium.
Yes you hear me right, after you add the mts reagent following drug treatment, you have to read the absorbance using a plate reader at a wavelength of 490nm. These factors may be cisacting, such as promoters and enhancers, or transacting, such as various dnabinding factors. The dualglo luciferase assay system is designed to work in growth media commonly used for mammalian cells with or without added serum. Usa toll free in usa 8003569526 6082744330 fax 6082772516. Promega corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, highvalue products for the life sciences. The conversion of mts into aqueous, soluble formazan is accomplished by dehydrogenase enzymes found in metabolically active cells. Celltiter 96 and 3hthymidine assays of hgmcsf using tf1 cells 21. Furthermore the mere presence of nms might influence the od measurement. A hc11 cells were seeded at a concentration of 10 4 per well into 96well microplates. The celltiter 96 aqueous one solution assay promega is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. Structures of mts tetrazolium and its formazan product. The mts, in the presence of phenazine methosulfate pms, is bioreduced by viable cells into a formazan product that is soluble in culture media fig.
Cell viability assays assay guidance manual ncbi bookshelf. The one solution contains mts compound and an electron coupling reagent pes. We used cell lysate obtained through 34 freezethaws of 1. Realtimeglo mt cell viability assay protocol promega.
Celltiter 96 aqueous nonradioactive cell proliferation. Shop online for a wide selection of promega celltiter 96 aqueous one solution cell proliferation assay mts homogeneous, colorimetric cell viability assay. Assay kit colorimetric ab197010 mts cell proliferation. The mts assay reagent is composed of mts and the electron coupling agent phenazine.
Cells were treated with or without compound saha or. Just as the manual states, i have found that lymphocyte cell lines. The active component is a novel tetrazolium compound called mts. Promega corpora on 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516 3. Promega corporation2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 6082744330 fax 6082772516. Jul 16, 2015 mtt assay principle analysis this lecture explains about the mtt assay principle. Choosing the right cellbased assay for your research. The tricky part here is to read the plate at the correct time. Briefly, at 3 h before each of the desired time points, 10 l of the mts reagent was added into each well and cells were incubated at 37 c for 3 h. Trypan blue exclusion showed less than 1% cell death both before and after the assays. Add 20 l mts solution containing pes to each well final concentration of mts will be 0. The conversion is thought to be carried out by nadphdependent dehydrogenase enzymes in metabolically active cells and the formazan dye produced can be quantified by measuring the absorbance at 490500 nm cory et al. Dulbeccos phosphatebuffered saline dpbs was added to each well of a 96well microplate with b 2.
Preparation of mts solution from celltiter 96 aqueous. This video lecture explains the importance of mtt assay and the mechanism of mtt assay. Certificate of analysis promega corporation 2800 woods hollow road madison, wi 537115399 usa telephone 6082744330 toll free 8003569526 fax 6082772516 internet. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516 3. This product is available through the promega helix onsite stocking program.
To help those who need to determine cell viability, promega has released the. Mts cell viability assay of upcyte hepatocyte cultures. A simple highcontent cell cycle assay reveals frequent. Aqueous one solution cell proliferation assay mts from promega. Celltiter 96 aqueous one solution cell proliferation assay. 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.
The pgl2 vectors carry the coding region for firefly photinus pyralis luciferase, used to monitor transcriptional activity in transfected eukaryotic cells. Promega celltiter 96 aqueous one solution cell proliferation. Aside from this, very little about the assay requires optimization, although the manual includes information on specific parameters that. Celltiter 96r aqueous one solution cell proliferation. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516 5. For an overview and information on the development of the dualglo luciferase assay system, see sections 6. Mtt assay principle analysis this lecture explains about the mtt assay principle. Both cell free controls cannot be calculated against values from cellular measurements. Celltiter 96 nonradioactive cell proliferation assay protocol. Theoretically, the color intensity of the formazan dye is correlated to the number of viable cells. Effect of b9 hybridoma cell number on color formation. 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.
Mts cell proliferation colorimetric assay kit k300. In vitro cell free reduction of mts the methodology for the in vitro cell free mts assay was essentially the same as that described for cells, with the following modifications. 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. The mtt cell proliferation assay is a colorimetric assay system that measures the reduction of a tetrazolium component mtt into an insoluble formazan product by the mitochondria of viable cells. Feb 24, 2010 mts assay posted in tissue and cell culture. Ive ordered dhtd073,bicalutamidephr1678 from sigma and enzalutamides1250 from selleck for mts assay which is to assess the proliferation of lncap,du145,22rv1 and pc3. Instructions for use of products g9711, g9712, g97. Mts is bioreduced by cells into a colored formazan product that is soluble in tissue culture medium. After incubation of the cells with the mtt reagent, a detergent solution is added to lyse the cells and solubilise the colour crystals. Biovisions mts cell proliferation assay kit is a colorimetric method for sensitive quantification of viable cells in proliferation and cytotoxicity assay. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. The mts is an alternative to mtt and the formazan formed from mts is watersoluble and less toxic. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526. The gel shift assay system contains target oligonucleotides, a control extract containing dnabinding proteins, binding buffer and reagents for phosphorylating oligonucleotides.
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. To assess the number of viable cells in culture several methods are available. Briefly this involves mixing 1 ml pms solution per 20 ml mts solution. A protocol for a simple and rapid method for detecting dnabinding proteins. Celltiter 96 aq ueous nonradioactive cell proliferation assay. To see articles that cite the use of the dualglo luciferase assay system, visit. 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. Prepare cells and test compounds in 96well plates containing a final volume of 100 lwell. Absorbance was measured at 490 nm on a spectramax plus384 reader molecular devices.
Promega corporation is a leader in providing innovative solutions and technical support to the life sciences industry. An optional set of wells can be prepared with medium only for background subtraction. Mts cell viability assay of upcyte hepatocyte cultures grown. This is a reliable system for obtaining experience with gel shift assays because ap2 binding activity is stable and produces a strong gel shift. Celltiter 96r aqueous one solution cell proliferation assay. Toll free in usa 8003569526 phone 6082744330 fax 6082772516. 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. Apr 16, 2010 for the mts assay, the celltiter 96 aq ueous one solution cell proliferation assay kit was used following the manufacturers instruction. The gel shift assay core system includes sufficient hela nuclear extract to perform 20 control reactions, gel shift binding 5x buffer, an sp1 consensus oligo and an ap2 consensus oligo. The assay is based on the cellular conversion of a tetrazolium salt into a formazan product that is easily detected using a 96well plate reader. Cells were treated with or without compound saha or staurosporine for 72 hours. 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.
This system can also be used to generate stable cell lines for cellbased assays to identify modulators of a. Celltiter 96 nonradioactive cell proliferation assay. Nonradioactive cell proliferation assay instructions for use of products g5421, g5430, g5440, g1111 and g1112. The absorbance of the formazan at 490nm can be measured directly from 96well assay plates without additional processing. Cell proliferation assay an overview sciencedirect topics. However, some chemicals or phytochemicals may change the activity of succinate dehydrogenase or interact with mtt directly. Pes has enhanced chemical stability, which allows it to be combined with mts to form a stable solution. Mts cell proliferation assay kit colorimetric 197010 is a colorimetric sensitive quantification of viable cells in proliferation and cytotoxicity assay. Standard dna purification protocol this section details our standard protocol for lysis of bacterial cell cultures and dna purification.
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