Seahorse xf96 glycolysis analyzer

Manufactured by Agilent Technologies

Sourced in United States

The Seahorse XF96 Glycolysis Analyzer is a laboratory equipment designed to measure the extracellular acidification rate (ECAR) of cells, which is an indicator of glycolytic activity. The instrument uses a 96-well plate format to simultaneously analyze multiple cell samples and provide real-time data on their glycolytic metabolism.

Automatically generated - may contain errors

Lab products found in correlation

Lactate assay kit wst, by Dojindo Laboratories (3 mentions) Glucose uptake cell based assay kit, by Cayman Chemical (2 mentions) Glucose uptake assay kit, by Abcam (1 mentions) Atp assay kit, by Beyotime (1 mentions) Atp assay kit, by Abcam (1 mentions) Seahorse xf cell mito stress test kit, by Agilent Technologies (1 mentions) Seahorse xf glycolysis stress test kit, by Agilent Technologies (1 mentions) Seahorse xf96 wave software, by Agilent Technologies (1 mentions)

Spelling variants of this product

Seahorse XF96 analyzer (215 citations) XF96 analyzer (164 citations) Seahorse XF96 (83 citations) Seahorse Analyzer (37 citations)

11 protocols using seahorse xf96 glycolysis analyzer

Glycolysis and Mitochondrial Stress Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) analyses were conducted to evaluate the effect of PVT1 depletion on glycolysis stress and cell mitochondrial stress using the Seahorse XF96 Glycolysis Analyzer (Seahorse Bioscience, MA, USA). For ECAR analysis, glucose, oligomycin, and 2-deoxyglucose were sequentially added in special medium. Glucose was first injected into the medium and catabolized to lactate and ATP with a corresponding increased ECAR value. Then, oligomycin was injected, which inhibited mitochondrial ATP production and shifted the energy production to glycolysis, with the corresponding increase in ECAR. The ECAR was reported in milli-pH (mpH) units per minute.
For OCR analysis, first the ATP synthase inhibitor oligomycin was injected into the medium, and the induced decrease in OCR was associated with the proton current resulting from ATP synthase. Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) was then injected, leading to rapid oxygen consumption. Compared with basal respiration, the induced increased respiratory capacity indicated the spare respiratory capacity. Finally, rotenone and antimycin A, electron transport chain inhibitors, were injected. Residual respiration corresponds to the non-mitochondrial respiration. The OCR was reported in units of picomoles per minute.

Chen J., Yu Y., Li H., Hu Q., Chen X., He Y., Xue C., Ren F., Ren Z., Li J., Liu L., Duan Z., Cui G, & Sun R. (2019). Long non-coding RNA PVT1 promotes tumor progression by regulating the miR-143/HK2 axis in gallbladder cancer. Molecular Cancer, 18, 33.

+ Open protocol

+ Expand

Glycolysis Stress Assessment via PIM1 Inhibition

Check if the same lab product or an alternative is used in the 5 most similar protocols

The procedure for measurements of the extracellular acidification rates (ECARs) was performed as described previously.19 (link) Briefly, we assessed the effect of PIM1 inhibition on glycolysis stress using the Seahorse XF96 Glycolysis Analyzer (Seahorse Bioscience, North Billerica, MA, USA).

Xue C., He Y., Hu Q., Yu Y., Chen X., Chen J., Ren F., Li J., Ren Z., Cui G, & Sun R. (2018). Downregulation of PIM1 regulates glycolysis and suppresses tumor progression in gallbladder cancer. Cancer Management and Research, 10, 5101-5112.

+ Open protocol

+ Expand

Quantifying Cellular Metabolism Using Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

The Glucose Uptake Assay Kit (abcam, USA), ATP Assay Kit (Beyotime, Shanghai, China), and Lactate Assay Kit-WST (Dojindo, Shanghai, China) were performed to assess the glycolysis level according to the manufacturer’s guidelines. Using the Seahorse XF96 Glycolysis Analyzer (Seahorse Bioscience, MA, USA), the ECAR and OCAR were measured according to the manufacturer’s guidelines.

Lin J., Wang X., Zhai S., Shi M., Peng C., Deng X., Fu D., Wang J, & Shen B. (2022). Hypoxia-induced exosomal circPDK1 promotes pancreatic cancer glycolysis via c-myc activation by modulating miR-628-3p/BPTF axis and degrading BIN1. Journal of Hematology & Oncology, 15, 128.

+ Open protocol

+ Expand

Quantifying TNBC Glycolysis Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols

Glycolysis of TNBC cells was evaluated by measuring the extracellular acidification rate (ECAR) using the Seahorse XF96 Glycolysis Analyzer (Seahorse Bioscience, MA, USA), as previously described.

Han G., Bai X., Li F., Huang L., Hao Y., Li W., Bu P., Zhang H., Liu X, & Xie J. (2023). Long non-coding RNA HANR modulates the glucose metabolism of triple negative breast cancer via stabilizing hexokinase 2. Heliyon, 10(1), e23827.

+ Open protocol

+ Expand

Glycolysis and Mitochondrial Stress in Pancreatic Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols

Glucose consumption, which was detected using the Glucose Uptake Cell-Based Assay Kit (Cayman, USA), and lactate production, which was measured using the Lactate Assay Kit-WST (Dojindo, Shanghai, China), were also quantified to detect the glycolysis level in A549 and NCI-H1299 cells. ATP synthesis was measured by fluorometric detection using the ATP Assay Kit (Ex/Em = 535/587 nm) (Abcam), according to the manufacturer's instruction. In addition, oxygen consumption rates (OCR) and extracellular acidification rates (ECAR) were considered as parameters to evaluate glycolysis stress and cell mitochondrial stress in pancreatic cancer cells using the Seahorse XF96 Glycolysis Analyzer (Seahorse Bioscience, MA, USA).

Cai Y., Zhao J., Luo C., Fang M., Yi Y., Chen Y., Huang P., Liao L, & Huang L. (2024). CD52 knockdown inhibits aerobic glycolysis and malignant behavior of NSCLC cells through AKT signaling pathway. Journal of Cancer, 15(11), 3394-3405.

+ Open protocol

+ Expand

Measuring Cellular Bioenergetics on Seahorse XF96

Check if the same lab product or an alternative is used in the 5 most similar protocols

For this assay, ECAR was measured on Seahorse XF96 Glycolysis Analyzer (Seahorse Bioscience, North Billerica, MA, USA) with Seahorse XF Glycolysis Stress Test Kit (Seahorse Bioscience) based on the user’s guidebook. In short, transfected cells were plated into a Seahorse XF 96 cell culture microplate, followed by the measurements of baseline. At the indicated time points, Glucose, Oligomycin (oxidative phosphorylation inhibitor) and 2-DG (glycolytic inhibitor) were sequentially injected. Similarly, OCR was detected with Seahorse XF Cell Mito Stress Test Kit (Seahorse Bioscience) on this Analyzer. Oligomycin, FCCP (reversible inhibitor of oxidative phosphorylation) and Rote + AA (mitochondrial complex I inhibitor rotenone plus the mitochondrial complex III inhibitor antimycin A) were injected. Finally, the results were analyzed with the Seahorse XF96 Wave software (Seahorse Bioscience).

Zhang X., Xu Y., Yamaguchi K., Hu J., Zhang L., Wang J., Tian J, & Chen W. (2020). Circular RNA circVAPA knockdown suppresses colorectal cancer cell growth process by regulating miR-125a/CREB5 axis. Cancer Cell International, 20, 103.

+ Open protocol

+ Expand

Evaluating Glycolytic and Mitochondrial Stress in Pancreatic Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols

The ECAR and OCR were considered as parameters to evaluate glycolysis stress and cell mitochondrial stress in pancreatic cancer cells using the Seahorse XF96 Glycolysis Analyzer (Seahorse Bioscience, MA, USA). In addition, glucose consumption, which was detected using the Glucose Uptake Cell-Based Assay Kit (Cayman, USA), and lactate production, which was measured using the Lactate Assay Kit-WST (Dojindo, Shanghai, China), were also quantified to detect the glycolysis level in pancreatic cancer cells.

Zhai S., Xu Z., Xie J., Zhang J., Wang X., Peng C., Li H., Chen H., Shen B, & Deng X. (2020). Epigenetic silencing of LncRNA LINC00261 promotes c-myc-mediated aerobic glycolysis by regulating miR-222-3p/HIPK2/ERK axis and sequestering IGF2BP1. Oncogene, 40(2), 277-291.

+ Open protocol

+ Expand

Extracellular Acidification Rate Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The extracellular acidification rate (ECAR) was measured using the Seahorse XF96 Analyzer Glycolysis (Seahorse Bioscience, USA), which calculates the net production and extrusion of protons into the extracellular medium. Treated and untreated cells were seeded in XF 96-well plates and incubated overnight at 37°C under 5% CO2 humidified atmosphere. Initially, the cells were incubated in the glycolysis stress test medium without glucose, and ECAR was assessed. Following this, D-glucose (10 mM), oligomycin (1 μM), and 2-deoxyglucose (100 mM) were injected in turn, and ECARs was assessed. The key parameters of glycolytic function, glycolysis and glycolytic capacity, were calculated by the XF Glycolysis Stress Test software.

Han D., Wei W., Chen X., Zhang Y., Wang Y., Zhang J., Wang X., Yu T., Hu Q., Liu N, & You Y. (2015). NF-κB/RelA-PKM2 mediates inhibition of glycolysis by fenofibrate in glioblastoma cells. Oncotarget, 6(28), 26119-26128.

+ Open protocol

+ Expand

Measuring Cellular Glycolytic Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols

The extracellular acidification rate (ECAR) was measured using a Seahorse XF96 Analyzer Glycolysis (Seahorse Bioscience, Santa Clara, USA) according to the manufacturer's protocol. Briefly, DLD-1 or RKO cells in 10% FBS RPMI-1640 were seeded in XF 96-well plates and incubated at 37°C in a 5% CO₂ humidified atmosphere overnight. The cells were then incubated in the glycolysis stress test medium without glucose, and the ECAR was measured. Following this, D-glucose (10 mM), oligomycin (1 μM), and 2-deoxyglucose (100 mM) were added into the wells at the indicated time points; meanwhile, corresponding ECARs were assessed. The ECAR values are presented as the mean ± SD of experimental triplicates. The key variables of glycolysis and glycolytic capacity were analyzed using XF Glycolysis Stress Test software.

Zuo S., Wu L., Wang Y, & Yuan X. (2020). Long Non-coding RNA MEG3 Activated by Vitamin D Suppresses Glycolysis in Colorectal Cancer via Promoting c-Myc Degradation. Frontiers in Oncology, 10, 274.

+ Open protocol

+ Expand

Metabolic Profiling of Colorectal Cancer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were determined by a Seahorse XF96 Analyzer Glycolysis according to the manufacturer’s instruction (Seahorse Bioscience). CRC cells were seeded in a XF96-well plate with the density of 10⁴ per well with REG1α overexpression or knockdown. As glycolysis occurs, the XF Analyzer can directly detect the acidification from the culture medium around CRC cells and reported as the ECAR. Following this, a saturating concentration of glucose, oligomycin and 2-deoxyglucose (2-DG) were added to the cells at the indicated time points. In the meanwhile, the ECARs were detected. For the assessment of OCR, oligomycin, Trifluoromethoxy carbonylcyanide phenylhydrazone (FCCP) and Rotenone were sequentially added to the cells.

Zhou M., He J., Li Y., Jiang L., Ran J., Wang C., Ju C., Du D., Xu X., Wang X., Li H., He F, & Wen H. (2023). N6-methyladenosine modification of REG1α facilitates colorectal cancer progression via β-catenin/MYC/LDHA axis mediated glycolytic reprogramming. Cell Death & Disease, 14(8), 557.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!