CD4+CD25− and CD4+CD25+ T cells were isolated using the mouse CD4+CD25+ T Cell Isolation Kit according to the manufacturer's instructions (Miltenyi Biotec). CD8+ T cells were isolated using the mouse CD8+ T Cell Isolation Kit according to the manufacturer's instructions (Miltenyi Biotec). For CD4+CD25+Treg suppression assay, CD4+CD25+ T cells were expanded in the presence of 10 ng/mL hIL‐2 and 10 μg/mL anti‐IFN‐γ with 0.25 μmol/L GO‐Y030 or 4 mmol/L L‐lactate (Cayman Chemical) for 3 days. CellTrace™ Violet Dye (Thermo Fisher Scientific)‐labeled naïve CD8+ T cells (0.8 × 105 cells) isolated from CD45.1 mice were cultured in a 96‐well plate with Dynabeads™ T‐activator CD3/CD28 (Veritask) in the presence or absence of CD4+CD25+ T cells.
L lactate
Manufactured by Cayman Chemical
L-lactate is a lab equipment product used for the detection and measurement of L-lactate in biological samples. It serves as a tool for researchers and scientists to quantify L-lactate levels, which is an important metabolite in various physiological processes.
Automatically generated - may contain errors
Lab products found in correlation
Mouse cd8 t cell isolation kit, by Miltenyi Biotec (1 mentions)
Celltrace violet dye, by Thermo Fisher Scientific (1 mentions)
Mouse cd4 cd25 t cell isolation kit, by Miltenyi Biotec (1 mentions)
Aconitase, by Cayman Chemical (1 mentions)
Xf96 cell culture microplate, by Agilent Technologies (1 mentions)
Xfe96 analyzer, by Agilent Technologies (1 mentions)
2 protocols using l lactate
1
Isolating and Expanding Regulatory T Cells
2
Mitochondrial and Glycolytic Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The Seahorse XFe96 Analyzer was used to measure mitochondrial respiration and glycolysis.
Briefly, BMDMs and PerMacs were plated at a density of 5x10 4 cells/well or 2x10 5 cells/ well in 80µL of culture medium in Agilent Seahorse XF96 Cell Culture Microplate. DMF (50 uM) and/or LPS (10 ng/ml) were added 3 hrs after the cells were plated. Following a 16 hr incubation, cells were washed and replaced with XF assay medium (Base Medium Minimal DMEM supplemented with 2 mM Ala-Gln, pH 7.4) prior to analysis. Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were measured after sequential addition of glucose 25 mM, oligomycin (1.5 uM), FCCP (1.5 µM)+ sodium pyruvate (1mM) and antimycin A (833 nM) and rotenone (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
(833 nM). L-Lactate (700510), Aconitase (705502) (Cayman); LDH (MK401) (TaKaRa) assay kits were commercially purchased. Standard protocols provided with the kits were followed when performing all assays.
Briefly, BMDMs and PerMacs were plated at a density of 5x10 4 cells/well or 2x10 5 cells/ well in 80µL of culture medium in Agilent Seahorse XF96 Cell Culture Microplate. DMF (50 uM) and/or LPS (10 ng/ml) were added 3 hrs after the cells were plated. Following a 16 hr incubation, cells were washed and replaced with XF assay medium (Base Medium Minimal DMEM supplemented with 2 mM Ala-Gln, pH 7.4) prior to analysis. Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were measured after sequential addition of glucose 25 mM, oligomycin (1.5 uM), FCCP (1.5 µM)+ sodium pyruvate (1mM) and antimycin A (833 nM) and rotenone (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
(833 nM). L-Lactate (700510), Aconitase (705502) (Cayman); LDH (MK401) (TaKaRa) assay kits were commercially purchased. Standard protocols provided with the kits were followed when performing all assays.
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