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Seahorse xf glycolytic rate assay kit

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The Seahorse XF Glycolytic Rate Assay Kit is a laboratory equipment product from Agilent Technologies. It is designed to measure the glycolytic rate in cells.

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Lab products found in correlation

Seahorse xf cell mito stress test kit, by Agilent Technologies (12 mentions) Seahorse xfe96 analyzer, by Agilent Technologies (10 mentions) Seahorse xf glycolysis stress test kit, by Agilent Technologies (3 mentions) Seahorse xfe24 analyzer, by Agilent Technologies (2 mentions) Seahorse xfe24 extracellular flux analyzer, by Agilent Technologies (2 mentions) Palmitate, by Merck Group (2 mentions) Oligomycin, by Agilent Technologies (2 mentions) Rotenone, by Agilent Technologies (2 mentions) 2 deoxyglucose, by Agilent Technologies (2 mentions) Seahorse xf96 cell culture microplate, by Agilent Technologies (2 mentions) Seahorse xf real time atp rate assay kit, by Agilent Technologies (2 mentions) Xf24 culture plate, by Agilent Technologies (2 mentions) Xf base medium, by Agilent Technologies (2 mentions) Mito stress test kit, by Agilent Technologies (2 mentions) Wave software, by Agilent Technologies (2 mentions) Seahorse xf24 extracellular flux analyser, by Agilent Technologies (1 mentions) Cell tak coated, by Corning (1 mentions) Poly d lysine, by Merck Group (1 mentions) Rapamycin, by Merck Group (1 mentions) Seahorse xf assay medium, by Agilent Technologies (1 mentions)

Spelling variants of this product

Glycolytic Rate Assay Kit (13 citations) XF Glycolytic Rate Assay Kit (12 citations) Glycolytic Rate Assay (9 citations) Agilent Seahorse XF Glycolytic Rate Assay Kit (8 citations) Seahorse XF Glycolytic Rate Assay (7 citations)

36 protocols using seahorse xf glycolytic rate assay kit

1

Seahorse XFe24 Glycolysis Assay

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Assays were performed using a Seahorse XFe24 analyzer (Seahorse Bioscience, Agilent Technologies) according to the Manufacturer’s instructions. The ECAR was measured using a Seahorse XF Glycolytic Rate Assay Kit (Seahorse Bioscience, Agilent, 103344-100). The glycolytic capacities of cells were analyzed with the Seahorse XF Glycolysis Rate/Stress Test Report Generator package. The %PER from glycolysis was calculated by subtracting the acidification from CO2 produced by the mitochondria, also called glycoPER.
Sun Q., Yuan M., Wang H., Zhang X., Zhang R., Wang H., Chen X., Zhu M., Liu S, & Wu J. (2021). PKM2 Is the Target of a Multi-Herb-Combined Decoction During the Inhibition of Gastric Cancer Progression. Frontiers in Oncology, 11, 767116.
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2

Glycolytic Profiling Using Seahorse Analyzer

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Assays were performed using a Seahorse XFe96 analyzer (Seahorse Bioscience, Agilent) according to the manufacturer’s instructions. The ECAR was measured using a Seahorse XF Glycolytic Rate Assay Kit (Seahorse Bioscience, Agilent, 103044-100) and a Seahorse XF Glycolysis Stress Test Kit (Seahorse Bioscience, Agilent, 103020-100). The glycolytic capacities of cells were analyzed with the Seahorse XF Glycolysis Rate/Stress Test Report Generator package. The %PER from glycolysis was calculated by subtracting the acidification from CO2 produced by the mitochondria.
Zhang Y., Zhao L., Yang S., Cen Y., Zhu T., Wang L., Xia L., Liu Y., Zou J., Xu J., Li Y., Cheng X., Lu W., Wang X, & Xie X. (2020). CircCDKN2B-AS1 interacts with IMP3 to stabilize hexokinase 2 mRNA and facilitate cervical squamous cell carcinoma aerobic glycolysis progression. Journal of Experimental & Clinical Cancer Research : CR, 39, 281.
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3

Assessing Cellular Bioenergetics Using Seahorse

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The ECAR and OCR were examined using Seahorse XF Glycolytic Rate Assay Kit (103344–100, Seahorse Bioscience, North Billerica, MA, USA) and Seahorse XF Mitochondrial Respiration Assay Kit (103260–100, Seahorse Bioscience) as per kits’ protocols, respectively and analyzed with the Seahorse XFe 24 Extracellular Flux Analyzer (Seahorse Bioscience) as previously described [15 (link)].
Zhao C., Wang B., Liu E, & Zhang Z. (2020). Loss of PTEN expression is associated with PI3K pathway-dependent metabolic reprogramming in hepatocellular carcinoma. Cell Communication and Signaling : CCS, 18, 131.
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4

Extracellular Flux Analysis of T Cells

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OCR and glycoPER were measured using a Seahorse XF24 Extracellular Flux Analyser (Seahorse Bioscience, Agilent Technologies, Santa Clara, CA). A Seahorse XF Cell Mito Stress Test Kit (103015-100; Seahorse Bioscience) and Seahorse XF Glycolytic Rate Assay Kit (103344-100; Seahorse Bioscience) were also used. T cells (3×105/well) were seeded in 24-well Cell-Tak-coated (22.4 μg/ml, Corning) Seahorse culture plates for analysis. For OCR measurement, 1.5 μM oligomycin, 0.5 μM FCCP and 0.5 μM antimycin-A/rotenone were added sequentially. To measure the glycoPER and mitoOCR/glycoPER ratio, 0.5 μM Rot/AA and 50 mM 2-DG were added sequentially.
Wen S., He L., Zhong Z., Zhao R., Weng S., Mi H, & Liu F. (2021). Stigmasterol Restores the Balance of Treg/Th17 Cells by Activating the Butyrate-PPARγ Axis in Colitis. Frontiers in Immunology, 12, 741934.
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5

Mitochondrial and Glycolytic Profiling

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1 × 104 cells were seeded into XFe96 cell culture plates in complete RPMI-1640 medium and incubated at 37 °C overnight in 5% CO2. To equilibrate temperature and pH of the detection system, cells were washed with assay RPMI-1640 medium and incubated at 37 °C for 1 h in a CO2-free incubator before assessment. OCR and ECAR were detected with an Agilent Seahorse XFe96 extracellular flux analyzer (Agilent Technologies, USA). To examine mitochondrial respiratory activity, cells were treated with oligomycin (1.5 μM), FCCP (1 μM) and rotenone/antimycin A (0.5 μM) using a Seahorse XF Cell Mito Stress Test Kit. For assessment of glycolytic activity, cells were treated with glucose (100 mM), oligomycin (10 μM) and 2-deoxy-D-glucose (2-DG, 500 mM) using a Seahorse XF Glycolytic Rate Assay Kit in sequence. All experiments were done in seven replicas each time and data expressed as means with SEM.
Dong S., Liang S., Cheng Z., Zhang X., Luo L., Li L., Zhang W., Li S., Xu Q., Zhong M., Zhu J., Zhang G, & Hu S. (2022). ROS/PI3K/Akt and Wnt/β-catenin signalings activate HIF-1α-induced metabolic reprogramming to impart 5-fluorouracil resistance in colorectal cancer. Journal of Experimental & Clinical Cancer Research : CR, 41, 15.
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6

Extracellular Acidification Measurement

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Assays were performed using a Seahorse XFe96 analyzer (Seahorse Bioscience, Agilent) according to the manufacturer’s instructions. The ECAR was measured using a Seahorse XF Glycolytic Rate Assay Kit (Seahorse Bioscience, Agilent, 103344-100). The contribution of mitochondria/CO2 to extracellular acidification was calculated by measuring cell oxygen consumption at the same time, and the acidification contributed by CO2 was subtracted from the total proton outflow rate (PER).
Guo D., Jin J., Liu J., Wang Y., Li D, & He Y. (2022). Baicalein Inhibits the Progression and Promotes Radiosensitivity of Esophageal Squamous Cell Carcinoma by Targeting HIF-1A. Drug Design, Development and Therapy, 16, 2423-2436.
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7

Glycolytic Capacity Analysis Using Seahorse XF

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Assays were performed using the Seahorse XFe96 analyzer (Seahorse Bioscience, Agilent) according to the manufacturer's instructions. ECAR was measured using Seahorse XF Glycolytic Rate Assay Kit(Seahorse Bioscience, Agilent,103044-100) and Seahorse XF Glycolysis Stress Test Kit(Seahorse Bioscience, Agilent 103020-100). Glycolytic capacities cells were analyzed by the Seahorse XF Glycolysis Rate/ Stress Test Report Generator package. %PER from glycolysis was calculated by subtracting the acidi cation from CO2 produced by the mitochondria.
Zhang Y., Zhao L., Yang S., Cen Y., Zhu T., Wang L., Xia L., Liu Y., Zou J., Xu J., Li Y., Cheng X., Lu W., Wang X., & Xie X. (2020). CircCDKN2B-AS1 Interacts with IMP3 to Stabilize Hexokinase 2 mRNA and Facilitate Cervical Squamous Cell Carcinoma Aerobic Glycolysis Progression.
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8

Evaluating Glycolytic Capacity via Seahorse XFe96

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Assays were performed using a Seahorse XFe96 analyzer (Seahorse Bioscience, Agilent) according to the manufacturer's instructions. The ECAR was measured using a Seahorse XF Glycolytic Rate Assay Kit (Seahorse Bioscience, Agilent, 103044-100) and a Seahorse XF Glycolysis Stress Test Kit (Seahorse Bioscience, Agilent, 103020-100). The glycolytic capacities of cells were analyzed with the Seahorse XF Glycolysis Rate/Stress Test Report Generator package. The %PER from glycolysis was calculated by subtracting the acidification from CO 2 produced by the mitochondria.
Zhang Y., Zhao L., Yang S., Cen Y., Zhu T., Wang L., Xia L., Liu Y., Zou J., Xu J., Li Y., Cheng X., Lu W., Wang X., & Xie X. (2020). CircCDKN2B-AS1 interacts with IMP3 to stabilize Hexokinase 2 mRNA and facilitate cervical squamous cellcarcinoma aerobic glycolysis progression.
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9

Mitochondrial and Glycolytic Function Assay

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The oxygen consumption rate (OCR) was measured using a Seahorse XF Cell Mito Stress Test Kit (Agilent, DE, USA), and the extracellular acidification rate (ECAR) was detected using a Seahorse XF Glycolytic Rate Assay Kit (Agilent) in an Agilent Seahorse Extracellular Flux Analyzer (Agilent). Briefly, the cells (2 × 104) were seeded in Seahorse cell culture microplates and incubated at 37°C overnight. The cultured medium was changed to XF assay medium, and the plates were placed in an incubator at 37°C without CO2 for 1 h. The OCR was measured by the sequential injections of 1 μM oligomycin, 1 μM carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), and 1 μM rotenone plus antimycin A (mitochondrial stress test). The ECAR was measured by the sequential injection of 0.5 μM rotenone plus antimycin A and 50 mM 2-DG (glycolytic rate assay).
Gao Q.Y., Zhang H.F., Tao J., Chen Z.T., Liu C.Y., Liu W.H., Wu M.X., Yin W.Y., Gao G.H., Xie Y., Yang Y., Liu P.M., Wang J.F, & Chen Y.X. (2021). Mitochondrial Fission and Mitophagy Reciprocally Orchestrate Cardiac Fibroblasts Activation. Frontiers in Cell and Developmental Biology, 8, 629397.
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10

Metabolic Profiling of Engineered iPSC-NK Cells

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WT iPSC-NK cells and CISH−/− iPSC NK cells were incubated with low concentration of IL-15 (1 ng/ml) for 7 days with or without rapamycin (100 ng/ml) (Sigma, R8781). Cells were resuspended in Seahorse XF Assay Medium (Agilent Technologies) and seeded at 150,000 cells/well (96-well plate). Plate was pre-coated with Poly-D-Lysine (Sigma, P6407) over-night at 37 °C. The extracellular acidification rate and the oxygen consumption rate were measured (pmoles/min) in real time in an XFe96 analyzer using Seahorse XF Glycolytic Rate Assay Kit (Agilent, 103344–100) and Seahorse XF Cell Mito Stress Test Kit (Agilent, 103015–100). Basal glycolysis was measured before the addition of rotenone/antimycin A (0.5 μM) and glycolytic was elucidated by subtracting the rate of glycolysis before and after addition of 2-deoxy-D-glucose (2DG, 100 mM). Basal respiration was measured after addition of glucose, ATP-linked respiration was calculated from subtraction of basal respiration and respiration after addition of oligomycin (1 μM), and mitochondrial spare respiratory capacity (SRC) was measured after addition of carbonyl cyanide-4 (trifluoromethoxy)phenylhydrazone (FCCP, 1 μM).
Zhu H., Blum R.H., Bernareggi D., Ask E.H., Wu Z., Hoel H.J., Meng Z., Wu C., Guan K.L., Malmberg K.J, & Kaufman D.S. (2020). Metabolic reprograming via deletion of CISH in human iPSC-derived NK cells promotes in vivo persistence and enhances anti-tumor activity. Cell stem cell, 27(2), 224-237.e6.
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