Introduction about this assay caymans xtt cell proliferation assay provides a tool for studying induction and inhibition of cell proliferation in any in vitro model. In addition to verifying the expected molecular weight of a gene construct, the tnt. Email promega technical services if you have questions on use of this system. Cell proliferation kit i mtt colorimetric assay mtt based for the nonradioactive quantification of cell proliferation and viability cat. Nanoglo luciferase assay system technical manualpdf. 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 dualglo luciferase assay system is a homogeneous reagent system that enables fast and simple quantitation of a stable. Terminal deoxynucleotidyl transferase tdt dutp nickend labeling tunel assay has been designed to detect apoptotic cells that undergo extensive dna degradation during the late stages of apoptosis.
The mtt cell viability assay kit provides a convenient, sensitive, quantitative and reliable assay for determining the number of viable cells in a given culture. Apoptotic cells can be detected by terminal deoxynucleotidyl transferase tdtmediated dutp nick end labeling tunel. Oh dna ends using the terminal deoxynucleotidyl transferase, recombinant, enzyme rtdt. In collaboration with phoenix flow systems, we offer the apobrdu tunel assay kit which provides all the materials necessary to label and detect the dna strand breaks of apoptotic cells. Tunel as a test for sperm dna damage in the evaluation of. The luciferase assay system incorporates coenzyme a coa for improved kinetics, allowing greater enzymatic turnover and resulting in increased light intensity that is nearly constant for at least 1 minute. The goscript reverse transcription system is a convenient kit that includes a reverse transcriptase and an optimized set of reagents for efficient synthesis of firststrand cdna optimized in preparation for pcr amplification. Oneglo ex luciferase assay system technical manualpdf. To determine this, we reached out to laurence delauriere, a senior applications scientist at promegafrance, who had never previously. M galectin1 or dtt buffer control as described in the annexin v flow cytometry section for 5. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 6082744330 fax 6082772516 printed in usa. The tunel assay is most commonly used to detect cells undergoing apoptosis, which is a form of programmed cell death. Use the cytospin to affix treated cells to microscope slides, and fix the cell. The transwell migration assay is a classical technique that allows scientists to quantify cell movement.
Oh dna ends using the terminal deoxynucleotidyl transferase, recombinant, enzyme rtdt, which forms a polymeric tail using the principle of the tunel tdtmediated dutp nickend labeling. This homogeneous colorimetric assay is based on the conversion of a tetrazolium salt mtt, a pale yellow substrate, to formazan, a purple dye. Hemagglutination ha assay protocol the hemagglutination assay is a method for titering influenza viruses based on their ability to attach to molecules present on the surface of red blood cells. Necrosis is caused by the cells inability to maintain homeostasis and is characterized by loss of plasma membrane integrity, cell swelling and lysis, random degradation of dna, and lack of an energy requirement or macromolecular synthesis. Tunel is an acronym for terminal deoxynucleotidyl transferase biotindutp nick end labeling. Necrosis is caused by the cells inability to maintain homeostasis and is characterized by loss of plasma membrane integrity, cell swelling and lysis, random degradation of dna, and lack of. Designing and implementing a new assay can be a challenging process with many unexpected troubleshooting steps. Trypan blue staining is a simple way to evaluate cell membrane. Goscript reverse transcription system protocol promega. The mtp activity assay kit is a sensitive, homogeneous fluorometric assay useful for continuous measurement of mtp activity in cell lysate or tissue homogenate. When dna strands are cleaved or nicked by nucleases, a large number of 3hydroxyl ends are exposed. Furthermore, the clickit tunel assay allows multiplexing with surface and intracellular biomarker detection.
Oh dna ends using the enzyme terminal deoxynucleotidyl transferase, recombinant rtdt. Migration refers to a cells ability to move individually or in clusters. Dualluciferase reporter assay system technical manual pdf. Neutral red cell cytotoxicity assay is one of the common methods used to detect cell viability or drug cytotoxicity. A test for basal cytotoxicity for an in vitro validation study phase iii. The assay is based on the extracellular reduction of wst8 by nadh produced in the mitochondria resulting in a watersoluble formazan. Cell movements are made possible through precise restructuring of their cytoskeleton and migration usually occurs in. Excerpts and links may be used, provided that full and clear credit is given to promega corporation with appropriate and specific direction to the original content. Test method protocol for the nhk neutral red uptake. The tunel assay allows the in situ detection of apoptosis in tissue sections. The apobrdu tunel assay kit provides the materials necessary to label dna strand breaks for the detection of apoptotic cells, as well as fixed samples of positive and negative control cells for assessing assay performance.
The luciferase assay system is an extremely sensitive and rapid reagent for. Cell counting kit8 technical manual general information cell proliferation assay and cytotoxicity assay for 100, 500, 3000, and 0 tests cell counting kit8 cck8 allows very convenient assays by utilizing dojindos highly watersoluble tetrazolium salt. Luciferase assay system quick protocol fb026pdf 102 kb english. The deadend colorimetric tunel system is a modified tunel assay designed to provide simple, accurate and rapid detection of apoptotic cells in situ at the singlecell level. Deadend fluorometric tunel system tunel tdtmediated dutp nickend labeling. Apobrdu tunel assay kit, with alexa fluor 488 antibrdu. The luciferase assay system yields linear results over at least eight orders of magnitude. A fretbased high throughput screening assay to identify. The assay is based on the extracellular reduction of wst8 by nadh produced in the mitochondria resulting in a watersoluble formazan which dissolves directly into the culture medium. The stabilized luciferase and proprietary buffer system improve assay. The assay is based on the enzymatic cleavage of the tetrazolium salt wst1 to formazan by cellular.
The deadend colorimetric tunel system provides the reagents to perform terminal deoxynucleotidyl transferasemediated dutp nickend labeling tunel of fragmented nuclear dna to assess apoptosis in situ at the singlecell level in tissue sections or cultured cells. The deadend colorimetric tunel system endlabels the fragmented dna of apoptotic cells using a modified tunel t dtmediated d u tp n icke nd l abeling assay. Oh dna ends using terminal deoxynucleotidyl transferase tdt, which forms a polymeric tail using the principle of the tunel tdtmediated dutp nickend labeling assay. Introduction about this assay caymans wst1 cell proliferation assay provides a tool for studying induction and inhibition of cell proliferation in any in vitro model. A protocol for a fluorescent tunel assay that measures the fragmented dna of apoptotic cells by catalytically incorporating fluorescein12dutp at 3. Wst8 cell proliferation assay kit provides a tool for studying induction and inhibition of cell proliferation in any in vitro model. The nanoglo luciferase assay system provides a simple, singleaddition reagent that generates a. Mtt proliferation assay protocol university of san diego. Promega corporation 2800 woods hollow road madison, wi 537115399 usa. Detection of nanoluc luciferase secreted from mammalian cells. The deadend fluorometric tunel system measures the fragmented dna of apoptotic cells by catalytically incorporating fluorescein12dutp at 3. Caspaseglo 37 assay is a homogeneous luminescent assay that measures caspase37 activities. The deadend colorimetric tunel system measures nuclear dna fragmentation. Instructions for use of products j1250, j1255, j3011, j3015, j4011, j4015.
Glucose uptakeglotm assay tm467 promega corporation. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. Get cells out of freezer, thaw on ice for 5 minutes 2. Nanoglo luciferase assay system promega corporation. The 10 ml volume is sufficient for the evaluation of 960 assays in ten 96well plates or 192 assays in eight 24well plates. Dualglo luciferase assay system promega corporation. The tunel assay can be adapted for analysis by light microscopy. The assay is based on the extracellular reduction of xtt by nadh produced in the mitochondria via. Promega corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, highvalue products for the life sciences. The pd1pdl1 blockade bioassay is a bioluminescent cellbased assay that can be used to measure the potency and stability of antibodies and other biologics designed to block the pd1pdl1 interaction. Apoptosis is a transient process with appearance and disappearance of markers over. We present a procedure for the comet assay, a gel electrophoresisbased method that can be used to measure dna damage in individual eukaryotic cells. Promega offers the pgl4 series of firefly and renilla luciferase vectors designed for use with the dlr assay systems. Ubiquinone coenzyme q10 hplc assay 28 catalog number.
Deadend fluorometric tunel system promega corporation. L of mtt solution to all wells of 48 h cultured lymphocytes and incubate for 4 h at 37c. During this period, formazon crystals will be formed at the bottom of each well. Shop online for a wide selection of promega deadend colorimetric tunel system a modified tunel assay designed for simple, rapid detection of apoptotic cells in situ at the. Clickit tunel alexa fluor imaging assay protocol thermo. The clickit tunel alexa fluor imaging assay has been tested in hela, a549, and cho k1 cells with a variety of reagents that induce apoptosis including staurosporine figure 6. The assay couples glucose oxidation and nadh production with a bioluminescent nadh detection system 1,2 figure 1.
Celltiterglo one solution assay technical bulletinpdf. A viral suspension may agglutinate the red blood cells, thus preventing them from settling out of suspension. Test method protocol for the nhk neutral red uptake cytotoxicity assay phase iii validation study. Assay principle cell biolabs cytoselect wst1 cell proliferation assay reagent provides a colorimetric format for measuring and monitoring cell proliferation. The celltiter 96 nonradioactive cell proliferation assay is a collection of qualified reagents that provide a rapid and convenient method of determining viable cell number in proliferation, cytotoxicity 1,2, cell attachment 3,4, chemotaxis 5, and apoptosis 6 assays. Detection of apoptotic cells in tissue samples currently relies on the tunel assay. A total of 18 readings for both negative and the positive controls and 80 readings for the. Tunel positive cardiomyocytes show morphological features of apoptosis and the typical ladder pattern in dna. To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. They are also applied in the screening of cytotoxic agents and lymphocyte activation. P450glo screening systems technical bulletinpdf 42 kb english.
Deadend colorimetric tunel system protocol promega. Deadend fluorometric tunel system technical bulletin, tb235. Glutamateglo assay technical manual, tm495 promega. This homogeneous colorimetric assay is based on the conversion of a tetrazolium salt mtt, a pale yellow substrate, to. See our terms and conditions for additional information. The p450glo cyp3a4 assay system provides a homogeneous, luminescent. The deadend fluorometric tunel systema is designed for the specific.
The principle of this assay is based on the detection of viable cells via the uptake of the dye neutral red. Terminal deoxynucleotidyl transferase tdt is a templateindependent dna polymerase that is normally active in primitive lymphoid cells to increase receptor diversity. The assay couples glutamate oxidation and nadh production with a bioluminescent nadh detection system 1,2 figure 1. Jun 27, 2006 we present a procedure for the comet assay, a gel electrophoresisbased method that can be used to measure dna damage in individual eukaryotic cells. Incubate at room temperature for 1 hour or overnight at 4c 3. Ubiquinone coenzyme q hplc assay eagle biosciences.
The t cell activation bioassay nfat is a bioluminescent cellbased assay that overcomes the limitations of existing assays and can be used for the discovery and development of novel biologic and cell therapies aimed at inducing, strengthening andor engineering t cell responses. We wanted to know what major snags a scientist new to the nanobret assay would encounter. Celltiter 96 nonradioactive cell proliferation assay. Tunel staining allows for visualization and quantification of apoptotic cells. The deadend colorimetric tunel system measures nuclear dna fragmentation, an important biochemical indicator of apoptosis. Dualluciferase reporter assay system promega corporation.
November 4, 2003 test method protocol for the nhk neutral red uptake cytotoxicity test. Rogers ms, cryan lm, habeshian ka, bazinet l, caldwell tp, ackroyd pc, et al. Intended purpose the eagle biosciences ubiquinone coenzyme q 10 hplc assay kit is intended for the quantitative determination of ubiquinone coenzyme q10 in plasma, serum and edtablood. In order to determine the number of viable cells cell proliferation kit xtt employs 2,3bis2methoxy4nitro. Tunel as a test for sperm dna damage in the evaluation of male infertility. March 2016 1 kit for 2,500 tests store at 15 to 25c 1. Quick system is ideal for screening large numbers of constructs for either naturally occurring or deliberately engineered mutations. Applications cell proliferation assays are widely used in cell biology for the study of growth factors, cytokines or media components.
Apoptosis is an important biological process during development, and for maintaining tissue homeostasis. Apoptosis assay tunel staining apoptosis was determined using the terminal deoxynucleotidyl transferase mediated dutp nickend labeling tunel assay deadend colorimetric tunel system, promega. Pipette out the spent media along with suspension of cultured cells. Cell movements are made possible through precise restructuring of their cytoskeleton and migration usually occurs in response to stimuli that act as cues.
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