Multiple parameters are obtained in one assay, including: basal respiration, ATP production-coupled respiration, maximal and reserve capacities and non-mitochondrial respiration. The XF Glycolysis Stress Test Kit is the first Standard for measuring glycolytic capacity in cells. Multiple parameters are obtained in one assay, including: basal respiration, ATP production-coupled respiration, maximal and reserve capacities and non-mitochondrial respiration. Cell Characterization: The XFp Analyzer and the XF Real-Time ATP Rate Assay. Assay result data must be generated by the same Agilent Seahorse XF, XFe or XFp Analyzer. The Glycolysis Stress Test kit contains the materials sufficient for six full plate tests and is available in two packaging sizes. Site Preparation Checklist. All assays were performed according to manufacturer instructions, as briefly described below. The Seahorse XFp Glycolysis Stress Test Kit makes it easy to assess glycolytic function in cells. By using the Agilent Seahorse XF Cell Mito Stress Test Kit, the Agilent Seahorse XF Glycolysis Stress Test Kit, and the Agilent Seahorse XFp Cell Energy Phenotype Test Kit, researchers can obtain functional metabolic data Mitochondrial Stress Test (MST) and Glycolysis Stress Test (GST) experiments were completed using a Seahorse XF96 extracellular flux analyzer (Seahorse Biosciences, Billerica, MA, USA) according to previously reported methods with minor modification (Liu et al., 2019). . Measure T cell activation with the use of programmable stimulant injection ports. Item ID: SEA101135100 Brand: Seahorse. For more information, see Supplementary Information. This bioavailable, conjugated fatty acid substrate frees you from the tedious, time-consuming, and variable conjugation process. 103017-100 XFp Glycolysis Stress Test Kit 6 XFp FluxPak (contains cartridges, miniplates, and calibrant) Part # Description # Cartridges 103022-100 XFp FluxPak 12 . When the % of PER from Glycolysis for a selected group is less than 50% or less than 10%, the Report Generator will display a warning message in the legend on the Summary Printout tab (below the Normalize button). RNA Preparation and RNA Sequencing PC for XFp data analysis. Agilent Seahorse XFp Glycolysis Stress Test Kit User Guide 7 Glossary • Glycolysis: The process of converting glucose to pyruvate. This assay kit utilizes a set of inhibitors to reveal the cells' ability to switch oxidative pathways in meeting basal energetic demands and provides information regarding the relative contributions of glucose, glutamine and long chain fatty acid . For Research Use Only. Seahorse XFp Glycolysis Stress Test kit. Details of these installations must be provided in . Seahorse XF24 V7 PS Cell Culture Microplates, Pack of 10. The Seahorse XF Glycolytic Rate Assay provides a more precise measurement of extracellular acidification due to glycolysis by subtracting out mitochondrial sources of acidification, as well as reporting the data in standard units (pmol/min). b Aged vs. young glycolysis The XFp Cell Mito Stress Test (103010-100) and Glycolysis Stress Test kits (103017-100) were purchased from Agilent Technologies (Palo Ato, CA, USA). 103674-100 for . View Product. Seahorse XF24 V7 PS Cell Culture Microplates, Pack of 10. All the reagents and assay conditions were followed by manufacturer's instructions. 2.11. Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were analyzed using the Seahorse XFp analyzer and the Seahorse XF Cell Mito Stress Test kit in samples taken at days 8 and 12. Item ID: SEA103017100 Brand: Seahorse. Agilent Seahorse XFp Glycolysis Stress Test Kit . Colony Formation Assay No. Seahorse XFp Extracellular Flux Analyzer. No. The Seahorse XF Glycolysis Stress Test run on the Seahorse XFp Analyzer illustrated an increase in both glycolysis (measured by extracellular acidification rate, ECAR) and glycolytic reserve compared to quiescent cells. 3 103017-100 Seahorse XFp Glycolysis Stress Test Kit 1 329,00 26,32- 302,68 302,68 Store at ambient temperature 4 103275-100 Seahorse XFp Cell Energy Phenotyp Tst Kt 1 299,00 23,92- 275,08 275,08 Store at ambient temperature 5 103010-100 Seahorse XFp Cell Mito Stress Glycolysis was measured under basal condition and in response to glucose (10 mM), oligomycin (1.5 μM), and 2-deoxyglucose (50 mM) following manufacturer's instructions (Agilent Seahorse XFp Glycolysis Stress Test Kit). Billerica, MA, June 13, 2014 - Seahorse Bioscience, the world leader in metabolic analyzers and XF stress test kits for measuring cell metabolism, introduces the XFp Extracellular Flux Analyzer ? 103270-100; Agilent Techno-logies), and Substrate Oxidation Stress Tests (Cat. The XFp Cell Mito Stress Test and XFp Glycolysis Stress Test kits contain pre-measured, verified compounds for assessing the two major energy pathways in cells and are recommended for characterizing the metabolic phenotype of cells using the XFp Analyzer. On the Y-axis, extracellular acidification rate is the mpH change over 1-min intervals, which is then expressed relative to 100,000 monocytes to account for between-well cell number variation. On the Y-axis, extracellular acidification rate is the mpH change over 1-min intervals, which is then expressed relative to 100,000 monocytes to account for between-well cell number variation. OCR, a marker for mitochondrial respiration, and ECAR, a marker for glycolysis, were analyzed using a Seahorse XFp Cell Mito Stress Test Kit or a Seahorse XFp Real Time ATP Rate Assay (Agilent Technologies). The Gold Standard for Metabolic Assays The XF Cell Mito and Glycolysis stress test kits provide the gold standard in metabolic assays. Agilent Seahorse XF Glycolysis Stress Test Kit User Guide Agilent Technologies 1 Introduction Introduction 6 Glossary 8 The Agilent Seahorse XF Glycolysis Stress Test is the standard assay for measuring glycolytic function in cells. Agilent Seahorse XF technology allows for the simultaneous measurement of the OCR and the ECAR in live, intact cells. a Example glycolysis stress test assay showing calculated parameters. a Example glycolysis stress test assay showing calculated parameters. The extracellular acidification rate (ECAR) was measured using a Seahorse XFp Glycolysis Stress Test Kit (103020-100, Agilent) with a Seahorse XFp Analyzer (Agilent) according to the manufacturer's instructions. Seahorse assay. XF assays are performed in a Seahorse XFp miniplate in conjunction with an XFp sensor cartridge. Multiple parameters are obtained in this one assay including, basal respiration, ATP-linked respiration, maximal and reserve capacities, and non-mitochondrial respiration. Agilent Seahorse XF Cell Energy Phenotype Test Kit FAQs. Seeding Adherent Cells in Agilent Seahorse XFp Cell Culture Miniplates. BILLERICA, Mass., April 25, 2012 /PRNewswire/ -- Seahorse Bioscience, the leader in cellular bioenergetic measurements, announced the launch of its newest stress test kit at the American Association of Cancer Research (AACR) Annual Meeting 2012. This data highlights the importance of discerning the energy pathways utilized by the cell for survival under normal and stressed conditions. No. In vitro stimulations . Glycolysis is examined using Seahorse XF Glycolysis Stress Test Kit (Agilent Technologies, North Billerica, MA) as previously described (12). Seahorse XF Glycolysis Stress Test Kit User Guide Timex Product Manuals & Software Downloads Find your watch on the list below and click on the language link for your manual and/or software. View Product. If you are still unable to locate your user manual, you may email Timex Customer Service or call us toll-free at 1-800-448-4639. Briefly, Tregs (0.5 × 10 6 /well) were plated on Cell-Tak-coated Seahorse culture plate for 45 min. A, The time course and live measurement of OCR with injections of stressor . The extracellular acidification rate (ECAR) was measured using a Seahorse XFp Glycolysis Stress Test Kit (103020-100, Agilent) with a Seahorse XFp Analyzer (Agilent) according to the manufacturer's instructions. Lysates were salvaged and total protein was quantified using a bicinchoninic acid kit (G Biosciences, MO, USA). an affordable extension to the XF product platform. The cell OXPHOS and cell glycolysis levels of HepG2 and A549 with or without PPP1R14B-AS1 knockdown were measured using a Seahorse XFp analyzer with a Seahorse XFp Cell Mito Stress Test Kit and a Glycolysis Stress Test Kit in accordance with the manufacturers' instructions. Seahorse XFp Glycolytic Rate Assay Kit The XFp Glycolytic Rate Assay provides a precise measure of glycolysis in live cells, revealing transient responses and rapid metabolic switches not discernible in endpoint lactate assays. XF Stress Test Kits provide reagents in single-use aliquots for consistency and convenience. Item ID: SEA103017100 Brand: Seahorse. 16. Using the XF Base Medium 100 mLwith the XF Glycolysis Stress Test Kit: To use XF Base Medium with the XF Glycolysis Stress Test Kit, supplement the XF Base Medium 100 mL with 2 mM (final concentration) L-glutamine (see table above). 103270-100) Agilent Seahorse XF Palmitate-BSA FAO Substrate (Cat No. For directly measuring glycolytic function, we recommend using the Seahorse XF Glycolysis Stress Test. 4 The Seahorse XF Palmitate-BSA FAO Substrate is a preconjugated, quality-controlled, fatty acid substrate for in vitro studies of long chain fatty acid oxidation by intact cells. 10 Agilent Seahorse XFp Glycolysis Stress Test Kit User Guide Kit Information Kit Contents The Seahorse XF Glycolysis Stress Test Kit includes six foil pouches. The output of the Cell Energy Phenotype test gives you the orientation from the basal state to the stress state, with the Stress OCR (%) and Stress ECAR (%) output. Cells rely on two major pathways, mitochondrial . Overall User Guides. No. Seahorse Cell Metabolism Analyses. . This comprehensive, easy-to-use kit measures the three key parameters of glycolytic flux: Glycolysis, Glycolytic Capacity, and Glycolytic Reserve, revealing critical information not evident in endpoint measurements. Respiration rates were normalised to µg protein per well. The Seahorse XFp Mito Stress Test Kit (Agilent Technologies, Inc.) was applied to measure the OCR. Conditioned NK cells were washed twice and resuspended in Refer to the Seahorse XF Glycolytic Rate Assay for a measure of lactate-associated proton efflux. Seahorse XFp analysis using the Cell Mito Stress Test kit was performed with acute injection of DMSO or 50 mM MB. Determine the ability of cells to use the glycolytic pathway to meet energy demand with the Seahorse XF Glycolysis Stress Test Quickly determine dependency of cellular energy production on glucose, glutamine, or fatty acids with the Seahorse XFp Mito Fuel Flex Test Kitand report generator. By directly measuring the extracellular acidification rate, (ECAR), see Figure 1 on page 7. PC for XFp data analysis. This new, lower cost instrument will enable many more labs access to Seahorse's unique technology. Using the gold standard Seahorse XF assays, the XFp provides a compact, easy-to-use bench top instrument at an affordable price. Every pouch includes one tube of each of the following compounds: glucose, XFp Site Preparation Checklist . The Seahorse XF Cell Mito Stress Test kit is an optimized solution for assessing mitochondrial function. 103020-100 SEAHORSE XF GLYCOLYSIS STRESS TEXT KIT(RE-AGENTS) United States: Chennai Air Cargo: PCS: 1: 36,221: 36,221: Oct 15 2016: 38220019: SEAHORSE XFP CELL MITO STRESS TEST KIT 103010-100: United States: Chennai Air Cargo: PCS: 1: 12,962: 12,962: Oct 15 2016: 39269099: SEAHORSE XFP FLUXPAK 103022-100(LAB ACCESSORIES) Now every lab can have direct access to Seahorse XF assays in their own lab. . • Glycolysis Rate: Activated > Control • Glycolytic Capacity: Activated > Control Item ID: SEA101135100 Brand: Seahorse. 103672-100 for Seahorse XF Long Chain Fatty Acid Oxidation Stress Test Kit, Cat. Agilent Technologies, Inc. Seahorse XF Palmitate-BSA FAO Substrate. This covers instrument time, the Seahorse cartridge plate, the Seahorse microplate for cell seeding, Seahorse media, reagents for a standard Mitochondrial Stress Test (MST) or Glycolysis Stress Test (GST), and SPARC staff time to help design and conduct the experiment, and analyze the data with you. minimum of 1-2 Seahorse XFp Extracellular Flux Analyzer systems in India. . 102504-100) Combining data from different analyzers is not supported currently (example: XF96 and XFe96). View Product. Seahorse XFp Glycolysis Stress Test kit. Agilent Seahorse XF Mito Fuel Flex Test Kit FAQs. The Agilent Seahorse Cell Mito Stress Test kit is the widely recognized, well accepted standard assay for assessing mitochondrial function. It has a formulation as follows. 103673-100 for Seahorse XF Glucose/ Pyruvate Oxidation Stress Test, Cat. Seahorse Capture Screen Insert Tool 1 reusable device. b Aged vs. young glycolysis stress test. The XF Glycolysis Stress Test run on the the XFp Analyzer illustrated an increase in both glycolysis (measured . XF Stress Test Kits provide reagents in single-use aliquots for consistency and convenience. Glycolysis stress test. Seahorse XFp Cell Mito Stress Test Kit The Seahorse XFp Cell Mito Stress Test kit is an optimized solution for assessing mitochondrial function. The cell glycolytic stress is evaluated by directly measuring the ECAR using the Seahorse XFp glycolysis stress test assay. Seahorse Bioscience, the world leader in metabolic analyzers and XF stress test kits for measuring cell metabolism, introduces the XFp Extracellular Flux Analyzer, an affordable extension to the XF product platform. 103020-100 and Cat No. After treatment with G-Rh2 (35 and 45 µM) for 24 h, HeLa cells (1×10 4 /well) were inoculated onto XFp cell culture plates for 24 h. The cells were then washed with analysis medium (supplemented with 10 mM glucose, 2 mM glutamine and 1 mM sodium . Note that no glucose or pyruvate is added to the XF Glycolysis Stress Test medium. Agilent Seahorse Mito Stress Test Single-File and Mulit-File XF Report Generator FAQs. View Product. This new, lower cost instrument will enable many more labs access to Seahorse's unique technology. Assay kits and reagents for performing gold standard XF assays are available. Every pouch includes one tube of each of the following compounds: oligomycin, FCCP and a mix of The Agilent Seahorse XF Glycolysis Stress Test presents the measure of glycolysis as the ECAR rate reached by a given cell after the addition of saturating amounts of glucose. Seahorse XF Palmitate-BSA FAO Substrate. Seahorse Capture Screen Insert Tool 1 reusable device. The Seahorse XF Palmitate-BSA FAO Substrate is a preconjugated, quality-controlled, fatty acid substrate for in vitro studies of long chain fatty acid oxidation by intact cells. Quantify ATP production by Glycolysis and OXPHOS simultaneously with the Seahorse XF real-time ATP rate assay Generate a metabolic phenotype within one hour with the Seahorse XF Cell Energy Phenotype Test Measure mitochondrial function with the Seahorse XFp Cell Mito Stress Test Kit and report generator. Cells cultured on Seahorse XFp plates were infected with RV-16 and a mitochondrial stress test was performed. Terms and Conditions . Startup consumables including XFp Fluxpak, XFp Mito Stress test kit, XFp Glycolysis Stress Test Kit, Seajorse XFp Cell energy Phenotype Test kit, XF Base Medium, and XFp carrier tray. 103275-100; Agilent Technologies), XFp Mito Fuel Flex Test Kit (Cat. Seahorse XFp Extracellular Flux Analyzer. XFp Real-Time ATP Rate Assay Kit - Rhino Bio Seahorse XFp Real-Time ATP Rate Assay Kit The Agilent Seahorse XFp Real-Time ATP rate assay is the only assay that measures and quantifies the rate of ATP (adenosine triphosphate) production from glycolysis and mitochondria simultaneously using label-free technology in live cells, in real time. 103275-100 Seahorse XFp Cell Energy Phenotype Test Kit EA 12 pouches2 XFp 103010-100 Seahorse XFp Cell Mito Stress Test Kit EA 6 pouches2 XFp 103017-100 Seahorse XFp Glycolysis Stress Test Kit EA 6 pouches2 XFp 103270-100 Seahorse XFp Mito Fuel Flex Test Kit EA 6 pouches2 XFp 103346-100 Seahorse XFp Glycolytic Rate Assay Kit EA 6 pouches2 XFp Item ID: SEA100777004 Brand: Seahorse. Item ID: SEA100777004 Brand: Seahorse. Agilent Seahorse XF Glycolysis Stress Test Kit (Cat No. Each miniplate is formatted to be a column of a typical 96-well plate, as shown . The test determines the ECAR source by sequential injection of glucose, oligomycin and 2-deoxy-glucose (2-DG) into the cell containing wells during the assay. Kit (Cat. Once the cancer has spread, the survival rate is less than 15%. Migration Assay The XF Glycolysis Stress Test reveals increased glycolytic activity in SOD1- derived fibroblasts as compared to healthy controls. Quickly determine dependency of cellular energy production on glucose, glutamine, or fatty acids with the Seahorse XF HS Mini Mito Fuel Flex Test Kit and report generator. Fig. For analysis of C. parvum infection- and inhibitor-induced glycolytic responses, the Glycolysis Stress Test Kit ® (Agilent, Rathingen, Germany) was applied using a Seahorse XFp extracellular flux analyser ® (Agilent, Rathingen, Germany). BILLERICA, Mass., June 13, 2014 /PRNewswire/ -- Seahorse Bioscience, the world leader in metabolic analyzers and XF stress test kits for measuring cell metabolism, introduces the XFp Extracellular . . Oligomycin, FCCP, and rotenone/antimycin A were added at the times indicated. Kit Contents The Seahorse XFp Cell Mito Stress Test Kit includes six foil pouches that each contain reagents sufficient for a complete Seahorse XFp Cell Mito Stress Test in the Agilent Seahorse XFp Cell Culture Miniplate. Run on a Seahorse XFp Analyzer, this assay quantifies proton efflux specific to glycolysis. The study was conducted using the XF24 Analyzer. No. This bioavailable, conjugated fatty acid substrate frees you from the tedious, time-consuming, and variable conjugation process. (ECAR) was measured on a Seahorse XFp analyzer (Seahorse Bioscience) using the Seahorse XF glycolysis stress test kit. Mitochondria are compartments in the cell that produce energy, and they play an important role in supporting the rapid growth of . Seahorse XF Glycolysis stress assay. 103017-100) Agilent Seahorse XF Mito Fuel Flex Test Kit (Cat No. XFp cell mito-stress test kit, XFp glycolysis stress test kit, and XFp Real-Time ATP rate assay kit (Seahorse Bioscience) were used to detect OCR, ECAR, and ATP ratio, respectively. The XF Glycolysis Stress Test run on the the XFp Analyzer illustrated an increase in both glycolysis (measured . Cells were seeded in Seahorse XFp cell . 103017-100 Seahorse XFp Glycolysis Stress Test Kit 6 103275-100 Seahorse XFp Cell Energy Phenotype Test Kit 12 103270-100 Seahorse XFp Mito Fuel Flex Test Kit 6 102504-100 Seahorse XF Plasma Membrane Permeabilizer 60 102720-100 Seahorse XF Palmitate-BSA FAO Substrate 30 Agilent Seahorse XF HS Mini Analyzer Each pouch contains reagents sufficient for an assay in a full 96- or 24-well XF Cell Culture Microplate. glycolytic function. Even in cases where the cancer is diagnosed and treated early, cells can sometimes survive treatment and spread to other organs. No. 103260-100 and Cat No. Seahorse experiments were performed on sorted cells of two donors using the Seahorse XFp Cell Mito Stress Test Kit and the XFp Glycolysis Stress Test Kit (Agilent Technologies, California, USA). The Seahorse XFp Mito Fuel Flex Test Kit provides an analytical method for measuring basal state mitochondrial fuel oxidation in live cells. The ECAR was detected using the Seahorse XF Glycolysis Stress Test Kit (Agilent Technologies, Inc.) on the Seahorse XFp Extracellular Flux Analyzer (Agilent Technologies, Inc.) according to the manufacturer's protocol. 102720-100) Agilent Seahorse XF Plasma Membrane Permeabilizer (Cat No. Colorectal cancer is the second largest cause of cancer deaths worldwide. The XF kit is for use with Agilent Seahorse XF96/XFe96, XFe24 Analyzers and the XFp kit is for use with the Agilent Seahorse XFp Analyzers. 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