ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit

Details for Product No. ABIN411718
Antigen
  • ATP
  • ATPIB
  • CAMRQ4
  • IB
  • ML-1
  • AI415030
  • Atpc1b
  • Ib
  • ATPase phospholipid transporting 8A2
  • ATPase, aminophospholipid transporter-like, class I, type 8A, member 2
  • ATP8A2
  • Atp8a2
Reactivity
Mammalian
4
3
2
2
2
2
2
2
2
2
2
1
1
1
Detection Method
Luminometric
Application
Cell Viability Assay (CVA), Detection (D)
Options
Brand ApoSENSOR™
Sample Type Cell Lysate, Plasma, Serum, Tissue Samples
Detection Method Luminometric
Specificity The ApoSENSOR™ Cell Viability Assay Kit utilizes bioluminescent detection of the ATP levels for a rapid screening of apoptosis and cell proliferation simultaneously in mammalian cells. The assay utilizes luciferase to catalyze the formation of light from ATP and luciferin, and the light can be measured using a luminometer or Beta Counter.
Characteristics ApoSENSORTM Cell Viability Assay Kit: Bioluminescent Assay for Rapid Screening of Apoptosis and Cell Proliferation Simultaneously in Mammalian Cells. Simple & Convenient.
Components Nucleotide Releasing Buffer
ATP Monitoring Enzyme
Enzyme Reconstitution Buffer
ATP (MW 551)
Target Name (Antigen)
Alternative Name ATP
Background Cell death (especially apoptosis) is an energy-dependent process that requires ATP. As ATP levels fall to a point where the cell can no longer perform basic metabolic functions, the cell will die. A typical apoptotic cell exhibits a significant decrease in ATP level. Therefore, loss of ATP level in cell has been used as an indicator of cell death. In contrast, cell proliferation has been recognized by increased levels of ATP. The ApoSENSOR™ Cell Viability Assay Kit utilizes bioluminescent detection of the ATP levels for a rapid screening of apoptosis and cell proliferation simultaneously in mammalian cells. The assay utilizes luciferase to catalyze the formation of light from ATP and luciferin, and the light can be measured using a luminometer or Beta Counter. The assay can be fully automatic for high throughput (10 seconds/sample) and is extremely sensitive (detects 10-100 mammalian cells/well). The high sensitivity of this assay has led to many other applications for detecting ATP production in various enzymatic reactions, as well as for detecting low level bacterial contamination in samples such as blood, milk, urine, soil, and sludge.
Application Notes Bioluminescent detection of the ATP level via luciferase catalyzed reaction for a rapid screening of apoptosis and cell viability in mammalian cells.
Comment

Further details regarding sample type: Cell and tissue lysates, culture media, urine, soil, sludge, plasma and serum, as well as many other biological fluids

Protocol A. Reagent Reconstitution and General Consideration: Reconstitute ATP Monitoring Enzyme with 220 µL/vial of the Enzyme Reconstitution Buffer. Mix well by gentle pipeting. The results in a yellow-green milky-like solution (not clear solution). Aliquot enough enzyme (1 µL per assay) for the number of assays to be performed in each experiment and refreeze the remaining at -70 °C for future use. The reconstituted enzyme is stable for up to 6 months at -70 °C. Protect the ATP Monitoring Enzyme from light as much as possible. Prepare an ATP standard solution by dissolving the 1 mg ATP into 1 mL of H2O. The solution is stable for several weeks at -20 °C. The ApoSENSOR TM kit is significantly more sensitive than other methods used for cell viability assays. The method can detect as few as 10 cells, but as a general guide, we recommend using 1 x 10 3 -10 4 cells per assay. Because of the high sensitivity of the ATP assay, avoid contamination with ATP from exogenous biological sources, such as bacteria or fingerprints. Ensure that the Nucleotide Releasing Buffer is at room temperature before use. The optimal temperature is 22 °C. Keep ATP Monitoring Enzyme on ice during the assay. The assay can be performed using either a single tube or a white walled 96-well luminometer plate (100 µL/well culture volume is recommended). B. Sample
1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction.
2. For suspension cells, transfer 10 µL of the cultured cells (containing 10 3 - 10 4 cells) into luminometer plate. Add 100 µL of the Nuclear Releasing Reagent. For adherent cells, remove culture medium and treat cells (10 3 - 10 4 ) with 100 µL of Nuclear Releasing Reagent for 5 minutes at room temperature with gentle shaking.
3. Add 1 µL ATP Monitoring Enzyme into the cell lysate. Read the sample in 1 minute in a luminometer.
4. Fold-decrease (or increase in the case of cell proliferation) in ATP levels can be determined by comparing these results with the levels of uninduced control. Note: The assay can be analyzed using cuvet-based luminometers or Beta Counters. When Beta Counter is used it should be programmed in the ''out of coincidence'' (or Luminescence mode) for measurement. The entire assay can also be done directly in a 96-well plate. It can also be programmed automatically using instrumentation with injectors (When using injector the ATP Monitoring Enzyme can be diluted with the Nuclear Releasing Buffer at 1:50 for injector. Mix a solution to the ratio of 1 µL ATP monitoring enzyme: 49 µL of Nucleotide Releasing Buffer. Add 50 µL per injection). C. Standard Curve: If the absolute ATP amount in samples needs to be calculated, an ATP standard curve should be generated (using the ATP standard provided in the kit) together with the above assays. Add 10 µL of a series of dilutions of ATP (e.g., 1, 0.1, 0.001, 0.0001, 0.00001, 0.000001 mg/mL, etc. Also includes a 0 mg/mL sample to measure background luminescence) to luminometer plates, then add 100 µL of Nuclear Releasing Reagent and 1 µL of ATP Monitoring Enzyme. Read the samples in 1 minutes in a luminometer (as described above). The background luminescence should be subtracted from all readings. The amount of ATP in uninduced and induced experimental samples can then be calculated from the standard curve.
Restrictions For Research Use only
Storage -20 °C
Expiry Date 12 months
Supplier Images
Image no. 1 for ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit (ABIN411718) Figure: (a) ATP Standard Curve. (b) Stability of luciferase at room temperature from ...
Product cited in: Zeng, Richani, Sutton-McDowall, Ren, Smitz, Stokes, Gilchrist, Thompson: "Prematuration with cyclic adenosine monophosphate modulators alters cumulus cell and oocyte metabolism and enhances developmental competence of in vitro-matured mouse oocytes." in: Biology of reproduction, Vol. 91, Issue 2, pp. 47, 2014 (PubMed).

Alavian, Dworetzky, Bonanni, Zhang, Sacchetti, Li, Signore, Smith, Gribkoff, Jonas: "The mitochondrial complex V-associated large-conductance inner membrane current is regulated by cyclosporine and dexpramipexole." in: Molecular pharmacology, Vol. 87, Issue 1, pp. 1-8, 2014 (PubMed).

Chini, Guerrico, Nin, Camacho-Pereira, Escande, Barbosa, Chini: "Targeting of NAD metabolism in pancreatic cancer cells: potential novel therapy for pancreatic tumors." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 20, Issue 1, pp. 120-30, 2014 (PubMed).

Liu, Chhipa, Pooya, Wortman, Yachyshin, Chow, Kumar, Zhou, Sun, Quinn, McPherson, Warnick, Kendler, Giri, Poels, Norga, Viollet, Grabowski, Dasgupta: "Discrete mechanisms of mTOR and cell cycle regulation by AMPK agonists independent of AMPK." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, Issue 4, pp. E435-44, 2014 (PubMed).

Paczkowski, Schoolcraft, Krisher: "Fatty acid metabolism during maturation affects glucose uptake and is essential to oocyte competence." in: Reproduction (Cambridge, England), Vol. 148, Issue 4, pp. 429-39, 2014 (PubMed).

Wilk, Wyczechowska, Zapata, Dean, Mullinax, Marrero, Parsons, Peruzzi, Culicchia, Ochoa, Grabacka, Reiss: "Molecular mechanisms of fenofibrate-induced metabolic catastrophe and glioblastoma cell death." in: Molecular and cellular biology, Vol. 35, Issue 1, pp. 182-98, 2014 (PubMed).

Chung-Davidson, Priess, Yeh, Brant, Johnson, Li, Nanlohy, Bryan, Brown, Choi, Li: "A thermogenic secondary sexual character in male sea lamprey." in: The Journal of experimental biology, Vol. 216, Issue Pt 14, pp. 2702-12, 2013 (PubMed).

Zeng, Ren, Guzman, Wang, Sutton-McDowall, Ritter, De Vos, Smitz, Thompson, Gilchrist: "Heparin and cAMP modulators interact during pre-in vitro maturation to affect mouse and human oocyte meiosis and developmental competence." in: Human reproduction (Oxford, England), Vol. 28, Issue 6, pp. 1536-45, 2013 (PubMed).

Li, Yu, Gu, Ma, Pasqualini, Arap, Snyder, Sidman: "Tissue plasminogen activator regulates Purkinje neuron development and survival." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, Issue 26, pp. E2410-9, 2013 (PubMed).

Zhong, Wang, Guo, Sagare, Fernández, Bell, Barrett, Griffin, Freeman, Zlokovic: "Protein S protects neurons from excitotoxic injury by activating the TAM receptor Tyro3-phosphatidylinositol 3-kinase-Akt pathway through its sex hormone-binding globulin-like region." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 30, Issue 46, pp. 15521-34, 2010 (PubMed).

Janssen, Horn, Niemann, Daniel, Schulze-Osthoff, Fischer: "Inhibition of the ER Ca2+ pump forces multidrug-resistant cells deficient in Bak and Bax into necrosis." in: Journal of cell science, Vol. 122, Issue Pt 24, pp. 4481-91, 2009 (PubMed).

Munemasa, Kim, Ahn, Kwong, Caprioli, Piri: "Protective effect of thioredoxins 1 and 2 in retinal ganglion cells after optic nerve transection and oxidative stress." in: Investigative ophthalmology & visual science, Vol. 49, Issue 8, pp. 3535-43, 2008 (PubMed).

Varma, Cheng, Voisine, Hart, Stockwell: "Inhibitors of metabolism rescue cell death in Huntington's disease models." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, Issue 36, pp. 14525-30, 2007 (PubMed).

Burnett, Melander, Puckett, Son, Wells, Dervan, Gottesfeld: "DNA sequence-specific polyamides alleviate transcription inhibition associated with long GAA.TTC repeats in Friedreich's ataxia." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, Issue 31, pp. 11497-502, 2006 (PubMed).

Ohshima: "Apoptosis and necrosis in senescent human fibroblasts." in: Annals of the New York Academy of Sciences, Vol. 1067, pp. 228-34, 2006 (PubMed).

Finkelstein, Brown, Avidor, Takeuchi: "The role of price, sociodemographic factors, and health in the demand for bariatric surgery." in: The American journal of managed care, Vol. 11, Issue 10, pp. 630-7, 2005 (PubMed).

Takeuchi, Mizuno, Zhang, Wang, Kawanokuchi, Kuno, Suzumura: "Neuritic beading induced by activated microglia is an early feature of neuronal dysfunction toward neuronal death by inhibition of mitochondrial respiration and axonal transport." in: The Journal of biological chemistry, Vol. 280, Issue 11, pp. 10444-54, 2005 (PubMed).

Diemer, Allen, Hales, Hales: "Reactive oxygen disrupts mitochondria in MA-10 tumor Leydig cells and inhibits steroidogenic acute regulatory (StAR) protein and steroidogenesis." in: Endocrinology, Vol. 144, Issue 7, pp. 2882-91, 2003 (PubMed).

Did you look for something else?