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商品描述
- Description
- Additional Information
- Readable Documents
- Assay Principle
- Reviews
Key Benefits
- Safe – Non Radioactive Enzyme release assay.
- Versatile – Useful for measuring activity of T Cells, Primary Cells, NK, complement and other lytic agents.
- Assay can be run in serum supplemented media.
- Homogenous – One-step, no wash assay. Assay can be run in same plate as samples.
- FAST – Results in 3-5 minutes. Chromium 51 or europium release for measurement are time consuming. The inherent sensitivity of luciferase detection is enhanced by the amplification effect of enzyme turnover, which produces thousands, millions or billions of high – energy molecules for each molecule of the enzyme.
- Highly Sensitive – Can detect fewer than 500 cells/well in the presence of serum or as few as 10 cells/well in serum-free or heat-killed media.
- GAPDH: The fact that GAPDH is a natural component of cells, and does not need to be introduced into the cells in any manner, distinguishes this assay from all methods which require prelabelling of cells, transfection, transformation, or other methods of introducing proteins or other molecules into the target cells in order to generate a signal in a later step.
- Advantages for measurement of cell mediated or complement mediated cytolysis – It is usually desirable to use smaller numbers of TCells than are needed for the 51Cr – release assay, since excessive numbers of effector cells can increase the background signal. This is now possible due to the high sensitivity of aCella-Tox.
- ADCC / CMC Assays – A non radioactive alternative to 51Cr assays. Please click here for a direct comparison between the aCella-TOX and (51Cr) Chromium Release Methods
- HTS – Adaptable for High Throughput format
- Non-destructive assay allows monitoring of additional parameters.
Additional information
| Kit Size | 500, 1000 |
|---|---|
| Includes | No Plates, 5 Lumi Plates, 5 Lumi Plates + 5 Tissue Culture Plates |
GAPDH is an important enzyme in the glycolysis and gluconeogenesis pathways. This homotetrameric enzyme catalyzes the oxidative phosphorylation of D-glyceraldehyde-3-phosphate to 1,3-diphosphoglycerate in the presence of cofactor and inorganic phosphate. In the aCella-TOX reaction scheme the release of GAPDH is coupled to the activity of the enzyme 3-Phosphoglyceric Phosphokinase (PGK) to produce ATP. ATP is detected via the luciferase, luciferin Bioluminescence methodology. Further, aCella-TOX is a homogeneous cytotoxicity assay; alternatively in dual mode, aCella-TOX can measure cytotoxicity and cell viability in the same plate. Culture supernatants can also be removed from the original plate and assayed in a different plate, allowing kinetics runs to be set up. The assay is non-destructive, allowing the monitoring of additional parameters such as gene expression.
| Document Title |
| aCella-TOX v1_3 Protocol |
| aCella-TOX Datasheet |
| msds.aCella-TOX |
| Title | File | Link | Author(s) | Journal | Year; Edition:Pages |
| Heat shock enhances the expression of cytotoxic granule proteins and augments the activities of tumor-associated antigen-specific cytotoxic T lymphocytes. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3468674/ | Takahashi A, Torigoe T, Tamura Y, et al. | Cell Stress & Chaperones | 2012;17(6):757-763 | |
| IGF-1R peptide vaccines/mimics inhibit the growth of BxPC3 and JIMT-1 cancer cells and exhibit synergistic antitumor effects with HER-1 and HER-2 peptides. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4368154/ | Foy KC, Miller MJ, Overholser J, Donnelly SM, Nahta R, Kaumaya PT | Oncoimmunology | 2014;3(11):e956005 | |
| HER-3 peptide vaccines/mimics: Combined therapy with IGF-1R, HER-2, and HER-1 peptides induces synergistic antitumor effects against breast and pancreatic cancer cells. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4368151/ | Miller MJ, Foy KC, Overholser JP, Nahta R, Kaumaya PT | Oncoimmunology | 2014;3(11):e956012 | |
| Phase I Active Immunotherapy With Combination of Two Chimeric, Human Epidermal Growth Factor Receptor 2, B-Cell Epitopes Fused to a Promiscuous T-Cell Epitope in Patients With Metastatic and/or Recurrent Solid Tumors. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2773479/ | Kaumaya PTP, Foy KC, Garrett J, et al. | Journal of Clinical Oncology | 2009;27(31):5270-5277 | |
| Identification of Cellular Proteins Required for Replication of Human Immunodeficiency Virus Type 1. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3448097/ | Dziuba N, Ferguson MR, O"Brien WA, et al. | AIDS Research and Human Retroviruses | 2012;28(10):1329-1339 | |
| Insulin-Like Growth Factor-1 Receptor Signaling Increases the Invasive Potential of Human Epidermal Growth Factor Receptor 2-Overexpressing Breast Cancer Cells via Src-Focal Adhesion Kinase and Forkhead Box Protein M1. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4293451/ | Sanabria-Figueroa E, Donnelly SM, Foy KC, et al. | Pharmacology | 2015;87(2):150-161 | |
| Combination Treatment with HER-2 and VEGF Peptide Mimics Induces Potent Anti-tumor and Anti-angiogenic Responses in Vitro and in Vivo. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3075707/ | Foy KC, Liu Z, Phillips G, Miller M, Kaumaya PTP | The Journal of Biological Chemistry | 2011;286(15):13626-13637 | |
| Resistance to Cytarabine Induces the Up-regulation of NKG2D Ligands and Enhances Natural Killer Cell Lysis of Leukemic Cells. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2586691/ | Ogbomo H, Michaelis M, Klassert D, Doerr HW, Cinatl J. | Neoplasia (New York, NY) | 2008;10(12):1402-1410 | |
| Anti-Tumor Effects of Peptide Therapeutic and Peptide Vaccine Antibody Co-targeting HER-1 and HER-2 in Esophageal Cancer (EC) and HER-1 and IGF-1R in Triple-Negative Breast Cancer (TNBC). | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4586465/ | Overholser J, Ambegaokar KH, Eze SM, et al. | Disis ML (Nora), ed. Vaccines | 2015;3(3):519-543 | |
| Generation and preclinical characterization of an antibody specific for SEMA4D. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4966508/ | Fisher TL, Reilly CA, Winter LA, et al. | mAbs | 2016;8(1):150-162 | |
| A Human Anti-M2 Antibody Mediates Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) and Cytokine Secretion by Resting and Cytokine-Preactivated Natural Killer (NK) Cells. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4411161/ | Simhadri VR, Dimitrova M, Mariano JL, et al. | Reeves RK, ed. PLoS ONE | 2015;10(4):e0124677 | |
| Natural Cytotoxicity Receptor-Dependent Natural Killer Cytolytic activity Directed at Hepatitis C Virus (HCV) Is Associated With Liver Inflammation, African American Race, IL28B Genotype, and Response to Pegylated Interferon/Ribavirin Therapy in Chronic HCV Infection. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3997579/ | Meng Q, Rani MRS, Sugalski JM, et al. | The Journal of Infectious Diseases | 2014;209(10):1591-1601 | |
| Myxoma Virus Infection Promotes NK Lysis of Malignant Gliomas In Vitro and In Vivo. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3677932/ | Ogbomo H, Zemp FJ, Lun X, et al. | Ulasov I, ed. PLoS ONE | 2013;8(6):e66825 | |
| Targeting a Glioblastoma Cancer Stem Cell Population Defined by EGF Receptor Variant III. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5661963/ | Emlet DR, Gupta P, Holgado-Madruga M, et al. | Cancer research | 2014;74(4):1238-1249 | |
| Genetically Associated CD16+56− Natural Killer Cell Interferon (IFN)-αR Expression Regulates Signaling and Is Implicated in IFN-α-Induced Hepatitis C Virus Decline. | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3295604/ | Conry SJ, Meng Q, Hardy G, et al. | The Journal of Infectious Diseases | 2012;205(7):1131-1141 |
| Reference |
| Methods and compositions for coupled luminescent assays. United States Patent 6,811,990 Corey and Kinders, issued November 2, 2004. |
| Corey, M. J. and Kinders, R. J. (2005) "Coupled Luminescent Methods in Drug Discovery: 3-Min Assays for Cytotoxicity and Phosphatase Activity" Drug Discovery Handbook, Ed. Shayne Cox Gad, published by John Wiley & Sons, Inc., pp. 689-731 |
| Corey, M.J., et al., "A Very Sensitive Coupled Luminescent Assay for Cytoxicity and Complement-Mediated Lysis," Journal of Immunological Methods 207:43-51, 1997. |
| Corey, M. J., et al., Mechanistic Studies of the Effects of Anti-factor H Antibodies on Complement-mediated Lysis,” Journal of Biological Chemistry 275: 12917-12925, 2000. |
| Schafer, H., et al., "A Highly Sensitive Cytotoxicity Assay Based on the Release of Reporter Enzymes, From Stably Transfected Cell Lines," Journal of Immunological Methods 204:89-98, 1997. |
| Racher, LDH Assay, in Cell and tissue culture: Laboratory procedures in biotechnology, A. Doyle and J.B. Griffiths, Eds. 1998, John Wiley & Sons: Chichester, New York, Weinheim. p. 71-5 |
| Decker, T. and Lohmann-Matthes, M.L. (1988) A quick and simple method for the quantitation of lactate dehydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity. J. Immunol. Meth. 115, 61-9. |
| Korzeniewski, C. and Callewaert, D.M. (1983) An enzyme-release assay for natural cytotoxicity. J. Immunol. Meth.64, 313-20. |
| Crouch, S.P.M., et al., "The Use of ATP Bioluminescence as a Measure of Cell Proliferation and Cytotoxicity," Journal of Immunological Methods 160:81-88, 1993. |
| Henry Ogbomo, Anke Hahn, Janina Geiler, Martin Michaelis, Hans Wilhelm Doerr, Jindrich Cinatl Jr. NK sensitivity of Neuroblastoma cells determined by a highly sensitive coupled luminescent method;Biochemical and Biophysical Research Comunications 339 (2006) pp375-379. Click here to read the publication |
| Part# | Reagent | Temperature |
| Part # 6001 | 4X Enzyme Assay Reagent | -20C |
| Part # 3008 | 1X Enzyme Assay Diluent | 2-8C |
| Part # 6003 | Glyeraldehyde 3-Phosphate (G3P) | -20C |
| Part # 6002 | 50X Detection Reagent | -20C |
| Part # 3009 | 5.5X Detection Assay Diluent | -20C |
| Part # 3035 | Lytic Agent | 2-8C |
| N/A | 5 Lumi Plates (Catalog# CLATOX100-3L) | N/A |
| N/A | 5 Lumi Plates + 5 Tissue Culture Plates (Catalog# CLATOX100-3P) | N/A |
