The XF Pro Analyzer features better OCR precision at low rates, verified instrument performance and repeatability specifications, optimized temperature control, and is automation enabled. The Seahorse XF Assay Learning Center provides protocols, techniques and resources aligned to each step of the XF workflow to ensure optimal assay performance and results. INTRODUCTION. On the day of the Seahorse assay, GM was replaced with XF medium (phenol red-free DMEM pH = 7.4 supplied with 25 mM glucose, 1 mM sodium pyruvate, and 2 mM glutamine) and incubated in a CO 2 -free Seahorse incubator for XF-calibrant (Agilent, 100840) and cartridge submerged in ddH 2 O were maintained in CO 2-free Seahorse incubator O.N. Agilent Seahorse XF Pro Analyzers measure the oxygen consumption (OCR) and extracellular acidification rate (ECAR/PER) of live cells in a 96-well format. Ryu et al. Aggressive and metastatic cancers show enhanced metabolic plasticity1, but the precise underlying mechanisms of this remain unclear. BMDMs were seeded on XF96 plates at 8 10 4 cells/well one day prior to the XF assay. reveal that sustained CR in humans reduces adipose SPARC. Purity CD4 + and CD8 + T cells of control and cKO mice were used for mitochondrial stress test in Seahorse XF Base Medium (Agilent) added with 10 mM glucose, 2 mM l-glutamine, and 1 mM sodium pyruvate with XF Cell Mito Stress Kit (Agilent). Elevation of SPARC promotes interferon-response and inflammation, while reduction of SPARC in adipocytes protects against aging-related The two-step process produces a consistent and even monolayer of cells. Agilent Seahorse Wave desktop software enables assay design and data analysis on all Quick Reference Guides; English; 01 Sep 2021; 129.20 KB; Agilent Seahorse XF Instruments Selection Guide Evaluate and compare the Seahorse XF Analyzers. Aggressive and metastatic cancers show enhanced metabolic plasticity1, but the precise underlying mechanisms of this remain unclear. Mitochondria, which are generally considered the cellular powerhouse, are small subcellular organelles that generate most cellular energy in the form of adenosine triphosphate (ATP). High-throughput drug screening was conducted on 10/15 cell lines (Table S5K). Caloric restriction (CR) reduces inflammation and enhances longevity, but the identity of CR-mimetics that improve health span is unclear. The assay was run on Seahorse XFe96 Analyzer (Agilent). Flow Cytometry Agilent Seahorse XF Pro Analyzers measure the oxygen consumption (OCR) and extracellular acidification rate (ECAR/PER) of live cells in a 96-well format. This mapping is traditionally carried out in one of two ways: a phenotype-centric, forward genetic approach that reveals the genetic changes that drive a phenotype of interest or a gene-centric, reverse genetic Agilent Seahorse XFe24 Analyzers measure the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of live cells in a 24-well plate format. For cells subjected to Seahorse XF Glycolysis Stress Test Kit (103020-100, Agilent Technologies), glucose (10 mM), oligomycin (1 mM) and 2-DG (50 mM) were injected sequentially. The XF Pro Analyzer features better OCR precision at low rates, verified instrument performance and repeatability specifications, optimized temperature control, and is automation enabled. The assay was run on Seahorse XFe96 Analyzer (Agilent). Anti-programmed death-1 (PD-1) immunotherapy that aims to restore T cell activity in cancer patients frequently leads to immune-related adverse events such as colitis. We have now placed Twitpic in an archived state. Oxygen consumption rate (OCR) of Xenopus oocytes was measured using a Seahorse XFe96 Analyser (Agilent) with Seahorse Wave software (Agilent, v2.6). In this study, Wu, Hu, Han et al. Purity CD4 + and CD8 + T cells of control and cKO mice were used for mitochondrial stress test in Seahorse XF Base Medium (Agilent) added with 10 mM glucose, 2 mM l-glutamine, and 1 mM sodium pyruvate with XF Cell Mito Stress Kit (Agilent). T-cell exhaustion has been described in chronic viral infection in mice, 14,15 humans, 16,17 and, Mitochondrial and glycolysis stress tests were performed on a Seahorse XFe96 Analyzer using corresponding kits (Agilent, Santa Clara, CA). The XF Assay Learning Center has been recently updated to reference the new Seahorse XF Real-Time ATP Rate Assay for performing initial cell characterization. Mitochondrial stress test and glycolytic stress test were performed in an XFe96 extracellular flux analyzer. The XF Assay Learning Center has been recently updated to reference the new Seahorse XF Real-Time ATP Rate Assay for performing initial cell characterization. Inhibitors were pre-treated before SPARC treatment. 1,2 Over the last two decades, research focusing on mitochondria-dependent cellular signaling and cell death has flourished. For cells subjected to Seahorse XF Glycolysis Stress Test Kit (103020-100, Agilent Technologies), glucose (10 mM), oligomycin (1 mM) and 2-DG (50 mM) were injected sequentially. To this end, we identified protein arginine N-methyltransferase 5 (PRMT5), cyclin-dependent kinase 9 (CDK9), and dihydroorotate dehydrogenase (DHODH) as novel synthetic lethal partners with gilteritinib treatment in AML.Using both genetic and pharmacologic approaches, we recapitulated the coessential nature of these genes in combination with Read 1 was used for identification of the Illumina library barcode, 8 bp cell barcode and 6 bp UMI and read 2 was used to map to the reference transcriptome of Hg38 with STAR (Dobin et al., 2013). Elevation of SPARC promotes interferon-response and inflammation, while reduction of SPARC in adipocytes protects against aging-related For cells subjected to Seahorse XF Glycolysis Stress Test Kit (103020-100, Agilent Technologies), glucose (10 mM), oligomycin (1 mM) and 2-DG (50 mM) were injected sequentially. Elevation of SPARC promotes interferon-response and inflammation, while reduction of SPARC in adipocytes protects against aging-related XF-calibrant (Agilent, 100840) and cartridge submerged in ddH 2 O were maintained in CO 2-free Seahorse incubator O.N. >OCR and ECAR or PER are key High-throughput drug screening was conducted on 10/15 cell lines (Table S5K). The Agilent Seahorse XFe96 Analyzer measures energy metabolism of live cells in real time. Next, we performed proteogenomic analysis for 15 iCCA cell lines to match the proteomic stratification, finding that three cell lines were concordant with S1, 5 with S2, 4 with S3, and 3 with S4 (Tables S5I and S5J). A two-step seeding process is recommended when seeding Agilent Seahorse XF24 Cell Culture Microplates. All strains were used at an MOI of 10 unless specified otherwise. The XF Assay Learning Center has been recently updated to reference the new Seahorse XF Real-Time ATP Rate Assay for performing initial cell characterization. T-cellrecruiting bispecific antibodies comprise a novel immunotherapeutic platform for the treatment of hematological malignancies and are currently being investigated in clinical trials. Mapping the relationship between genetic changes and their phenotypic consequence is critical to understanding gene and cellular function. XF-calibrant (Agilent, 100840) and cartridge submerged in ddH 2 O were maintained in CO 2-free Seahorse incubator O.N. Dear Twitpic Community - thank you for all the wonderful photos you have taken over the years. Please consult the Seahorse Cell Reference Database and/or the XF Publication database to provide an initial starting point for cell density values specific to your needs. >OCR and ECAR or PER are key This 96-well metabolic analyzer reports rates of mitochondrial respiration, glycolysis, and ATP production for a systems-level view of cellular metabolic function. Anti-programmed death-1 (PD-1) immunotherapy that aims to restore T cell activity in cancer patients frequently leads to immune-related adverse events such as colitis. T-cell exhaustion has been described in chronic viral infection in mice, 14,15 humans, 16,17 and, Mitochondrial and glycolysis stress tests were performed on a Seahorse XFe96 Analyzer using corresponding kits (Agilent, Santa Clara, CA). 1. The Seahorse XF Assay Learning Center provides protocols, techniques and resources aligned to each step of the XF workflow to ensure optimal assay performance and results. Starting from frozen vials of 3060 10 6 splenocytes, this usually allowed us to sort between 20 and 200 cells of interest, with only a handful of donors above 100 cells. Inhibitors were pre-treated before SPARC treatment. 35 Mitochondrial defects or The assay was run on Seahorse XFe96 Analyzer (Agilent). Inhibitors were pre-treated before SPARC treatment. Caloric restriction (CR) reduces inflammation and enhances longevity, but the identity of CR-mimetics that improve health span is unclear. Ryu et al. 1 The CD20xCD3 bispecifics glofitamab 2,3 and mosunetuzumab 4 have shown encouraging results in phase-I trials in patients with indolent or aggressive lymphomas. The XF Pro Analyzer features better OCR precision at low rates, verified instrument performance and repeatability specifications, optimized temperature control, and is automation enabled. Flow Cytometry At day 5, MDA-MB-231 cells were reseeded into XFp Cell Culture Miniplates (Agilent, 103025-100) at density 8 10 3 cells per well. At day 5, MDA-MB-231 cells were reseeded into XFp Cell Culture Miniplates (Agilent, 103025-100) at density 8 10 3 cells per well. Agilent Seahorse XF Pro Analyzers measure the oxygen consumption (OCR) and extracellular acidification rate (ECAR/PER) of live cells in a 96-well format. Extracellular acidification occurs in inflamed tissue and the tumor microenvironment; however, a systematic study on how pH sensing contributes to tissue homeostasis is lacking. After 48 h in the presence of VB5, cells were seeded at 7,500 cells per well in an Agilent XF 96-well plate (Agilent, 101085-004). The accession number for the single cell sequencing reported in this paper is Gene Agilent: 102601-100: Seahorse XF base medium, sterile, 1 L, 2/pk: Agilent: 102353-100: Feotal calf serum (AIEC) reference strain LF82.30, Staphylococcus aureus (NCTC 6571). This 96-well metabolic analyzer reports rates of mitochondrial respiration, glycolysis, and ATP production for a systems-level view of cellular metabolic function. Cells were adhered to each well of the Seahorse plate (Seahorse/Agilent, USA) using Cell-Tak (Corning, USA). Agilent Seahorse Wave desktop software enables assay design and data analysis on all Quick Reference Guides; English; 01 Sep 2021; 129.20 KB; Agilent Seahorse XF Instruments Selection Guide Evaluate and compare the Seahorse XF Analyzers. Oxygen consumption rate (OCR) of Xenopus oocytes was measured using a Seahorse XFe96 Analyser (Agilent) with Seahorse Wave software (Agilent, v2.6). Please consult the Seahorse Cell Reference Database and/or the XF Publication database to provide an initial starting point for cell density values specific to your needs. The Seahorse XF Assay Learning Center provides protocols, techniques and resources aligned to each step of the XF workflow to ensure optimal assay performance and results. The Seahorse XF96 analyzer (Agilent Technologies) was used to measure the ECAR. Mitochondria, which are generally considered the cellular powerhouse, are small subcellular organelles that generate most cellular energy in the form of adenosine triphosphate (ATP). This mapping is traditionally carried out in one of two ways: a phenotype-centric, forward genetic approach that reveals the genetic changes that drive a phenotype of interest or a gene-centric, reverse genetic approach that Dear Twitpic Community - thank you for all the wonderful photos you have taken over the years. Extracellular acidification occurs in inflamed tissue and the tumor microenvironment; however, a systematic study on how pH sensing contributes to tissue homeostasis is lacking. High-throughput drug screening was conducted on 10/15 cell lines (Table S5K). The Seahorse XF96 analyzer (Agilent Technologies) was used to measure the ECAR. Purity CD4 + and CD8 + T cells of control and cKO mice were used for mitochondrial stress test in Seahorse XF Base Medium (Agilent) added with 10 mM glucose, 2 mM l-glutamine, and 1 mM sodium pyruvate with XF Cell Mito Stress Kit (Agilent). 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