Pyruvate

Manufactured by Corning

Sourced in United States

Pyruvate is a chemical compound that serves as an important intermediate in various metabolic pathways. It is a key component in cellular respiration and energy production processes. Pyruvate can be used as a substrate or precursor for a range of laboratory experiments and analyses.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) L glutamine, by Corning (3 mentions) Dulbecco modified eagle medium (dmem), by Corning (3 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions) Glutamine, by Corning (2 mentions) Hepes, by Corning (2 mentions) Complete rpmi, by Corning (2 mentions) Lipopolysaccharides (lps), by InvivoGen (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Corning (2 mentions) Sodium pyruvate, by Corning (1 mentions) Fbs dmem, by Corning (1 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) Gapdh activity assay kit, by Abcam (1 mentions) X tremegene transfection reagent, by Roche (1 mentions) Puromycin, by Merck Group (1 mentions) Gentamicin, by Lonza (1 mentions) Flexmap3d, by Bio-Rad (1 mentions) Bio plex manager, by Bio-Rad (1 mentions) Lps e coli o111 b4, by Merck Group (1 mentions)

Spelling variants of this product

Sodium pyruvate (327 citations) L-glutamine and sodium pyruvate (5 citations) Glucose and sodium pyruvate (5 citations) DMEM) without pyruvate (2 citations) L-glutamine & sodium pyruvate (2 citations) L-Glutamine 4.5 g/L glucose and sodium pyruvate (2 citations)

10 protocols using pyruvate

Pyruvate Supplementation Assay

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For pyruvate treatment studies, the medium from cells grown in 10% FBS-DMEM containing pyruvate (Corning) was aspirated, washed with PBS three times, and replaced with 10% FBS-DMEM without sodium pyruvate (Corning). Supplemental sodium pyruvate (Thermo Fisher) was added at specific concentrations to the appropriate wells.

Duan X., Li S., Holmes J.A., Tu Z., Li Y., Cai D., Liu X., Li W., Yang C., Jiao B., Schaefer E.A., Fusco D.N., Salloum S., Chen L., Lin W, & Chung R.T. (2018). MicroRNA 130a Regulates both Hepatitis C Virus and Hepatitis B Virus Replication through a Central Metabolic Pathway. Journal of Virology, 92(7), e02009-17.

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Analyzing Inflammasome Activation in T2DM

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Human peripheral blood mononuclear cells (huPBMCs) from normal control (ctrl) and type 2 diabetes mellitus (T2DM) patients were purchased from Precision for Medicine, inc (BuyPBMC). Cells were maintained in vapor phase of liquid nitrogen (−120°C). On the day of experiment, huPBMCs were thawed in 37°C water bath and gently pipetted into 10mL of complete RPMI (Corning) supplemented with 10% FBS (Atlanta Biologicals), 2mM glutamine (HyClone), 1mM HEPES (Corning) and 1mM pyruvate (Corning) and centrifuged at 500g for 10min. Media was aspirated and cells were resuspended in 1mL of complete RPMI with supplements. 100,000 cells were plated in each well of a 96 well plate and incubated overnight. Stimulation with LPS (Invivogen) was performed the next day. 200ng/ml of LPS was given to the cells for 6hrs with or without 1400W (50μM). Cells were then treated with 2mM ATP (Sigma) for 30min and media was collected for huIL-1β ELISA (DY201–05). Cellular GAPDH Activity was assessed using GAPDH activity assay kit (abcam, ab204732).

De Jesus A., Keyhani-Nejad F., Pusec C.M., Goodman L., Geier J.A., Stoolman J.S., Stanczyk P.J., Nguyen T.A., Xu K., Suresh K.V., Chen Y., Rodriguez A.E., Shapiro J.S., Chang H.C., Chen C., Shah K.P., Ben-Sahra I., Layden B.T., Chandel N.S., Weinberg S.E, & Ardehali H. (2022). Hexokinase 1 cellular localization regulates the metabolic fate of glucose. Molecular cell, 82(7), 1261-1277.e9.

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Establishing Syndecan-4 Knockdown in C2C12 Myoblasts

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C2C12 mouse myoblast cells (ATCC; Manassas, VA, United States) were cultured in high-glucose Dulbecco’s modified Eagle’s medium containing 4.5 g/L glucose, 584 mg/L glutamine and 110 mg/L pyruvate (Corning, NY, United States) supplemented with 65 μg/mL gentamicin (Lonza, Basel, Switzerland), and 20% fetal bovine serum (Gibco/Thermo Fisher Scientific, Waltham, MA, United States). To achieve syndecan-4 knockdown, C2C12 cells were transfected stably with plasmids expressing short hairpin RNAs (shRNAs) specific for mouse syndecan-4 (shSDC4#1 and shSDC4#2) or a scrambled target sequence. The plasmids were obtained from OriGene (TR513122; Rockville, MD, United States) and targeted the following sequences: 5’-GAA CTG GAA GAG AAT GAG GTC ATT CCT AA-3’ (shSDC4#1), 5’-GCG GCG TGG TAG GCA TCC TCT TTG CCG TT-3’ (shSDC4#2) and 5’-GCA CTA CCA GAG CTA ACT CAG ATA GTA CT-3’ (scrambled). X-tremeGENE transfection reagent (Roche, Basel, Switzerland) was used for the transfection procedures. Transfected cells were then selected in medium containing 4 μg/mL puromycin (Sigma-Aldrich, St. Louis, MO, United States).

Becsky D., Szabo K., Gyulai-Nagy S., Gajdos T., Bartos Z., Balind A., Dux L., Horvath P., Erdelyi M., Homolya L, & Keller-Pinter A. (2020). Syndecan-4 Modulates Cell Polarity and Migration by Influencing Centrosome Positioning and Intracellular Calcium Distribution. Frontiers in Cell and Developmental Biology, 8, 575227.

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Profiling Cytokine Responses in Activated PBMCs

Cited 1 time

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PBMCs were archived as published (21 (link)), then thawed at 37°C before dilution in RPMI1640 with 2mM
L-glutamine, 25mM HEPES, 1% 100X Penicillin/Streptomycin (Gibco), 10% FBS (all
from Gibco), and 1mM pyruvate (Corning). Cells were resuspended in RPMI-1640 at
10⁶ cells/mL, seeded into 24-well plates (CellStar) at 500,000
cells/well, then stimulated with E. Coli O111:B4 LPS (25 ng/mL,
Millipore Sigma) to primarily target myeloid cells or human
αCD3/αCD28 Dynabeads (1 bead/cell, Gibco) to primarily target T
cells, for 20-72 hours. Aliquots of supernatants were stored at
−80°C for analysis after ≤2 freeze/thaw cycles. Cytokines
were quantified using a 25-plex Th17 magnetic bead kit (Millipore Sigma) and a
Bio-Rad FLEXMAP 3D with Luminex xPONENT 4.2 and Bio-plex Manager (Bio-Rad)
softwares with 1:10 dilutions as appropriate. Values below the level of
detection were replaced with a value that was 1/10 of the minimum standard curve
value (specific to each cytokine) for analysis.

Pugh G.H., Fouladvand S., SantaCruz-Calvo S., Agrawal M., Zhang X.D., Chen J., Kern P.A, & Nikolajczyk B.S. (2022). T cells dominate peripheral inflammation in a cross-sectional analysis of obesity-associated diabetes. Obesity (Silver Spring, Md.), 30(10), 1983-1994.

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Lysosome-related Organelle Regulation

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Hank’s balanced saline solution (HBSS), pyruvate, and L-glutamine were purchased from Corning. Chloroquine (CQ) was purchased from MP Biomedicals. Bafilomycin A (baf A), leupeptin, and Torin 1 were purchased from Cayman chemical. Carbonyl cyanide m-chlorophenylhydrazone (CCCP) was purchased from Enzo Life Sciences. Bortezomib was purchased from LC Laboratories. Pepstatin, glycine, lysine and serine were purchased from Sigma. Glucose, asparagine, and histidine were purchased from Fisher. Arginine was purchased from Acros Organics. The siRNAs against TFEB, TFE3, and SNAT7(siGenome SMARTpool) and the control scramble siRNA were purchased from Dharmacon. Rabbit antibodies used in this study were against LC3B (Abcam), P62, S6K, pT389-S6K, ATG5, ATG7, TFEB, TFE3 (Cell Signaling Technology), SNAT7 (Sigma), and Limp2 (Novus Biologicals). Mouse antibodies used in this study were against GAPDH (Developmental Studies Hybridoma Bank (DSHB)), Lamp1 [38 (link)](DSHB), and CD63 [39 (link)] (DSHB). TFEB-EGFP was a gift from Shawn Ferguson (Addgene plasmid #38119) [10 (link)]. HRP-conjugated secondary antibodies for Western blotting were purchased from Jackson Laboratory. Secondary antibodies conjugated to Alexa Fluor were purchased from Molecular Probes.

Wilden A.R., Molina J.A., Feuerborn M., Boyle D, & Lee S.Y. (2018). Glutamine-dependent lysosome homeostatic changes induced by starvation and lysosome inhibition. Biochimica et biophysica acta. Molecular cell research, 1865(9), 1356-1367.

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Isolation and Stimulation of Peritoneal Macrophages

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Peritoneal macrophages were isolated from 8–10-week-old ΔE1HK1 and littermate control WT mice by peritoneal lavage as described previously (Ray and Dittel, 2010 ). Briefly, mice were euthanized and the peritoneal cavity was injected with PBS supplemented with 5% FBS using a 27g needle. The abdominal wall was gently agitated to dislodge peritoneal cells. Using a fresh 25g needle, the peritoneal lavage (PBS and peritoneal cells) was aspirated and collected. Cells were then centrifuged at 500g for 10min and re-suspended in 1mL of complete RPMI (Corning) supplemented with 10% FBS (Atlanta Biologicals), 2mM glutamine (HyClone), 1mM HEPES (Corning) and 1mM pyruvate (Corning). Stimulation with LPS (Invivogen) was performed the next day. 300ng/ml of LPS was given to the cells for 4–6hrs with or without the following drugs: OT (50μM), 6-AN (1mM), 1400W (50μM), or CGP3466 (50nM).

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Culturing Caco-2, Raw264.7, and HBEpiC Cells

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Caco-2 and Raw264.7 cell lines, purchased from Bioresource Collection and the Research Center (BCRB; Hsinchu, Taiwan), were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Corning, NY, USA) containing 10% inactive Fetal Bovine Serum (FBS, Corning) and 1 mM pyruvate (Corning). Caco-2 cells were supplemented with 0.01 mg/mL human transferrin (Sigma-Aldrich Chemical Co.). The human bronchial epithelial (HBEpiC, Cat:3210) cell line was purchased from ScienCell (Carlsbad, CA, USA) and maintained in Bronchial Epithelial Cell Medium (BEpiCM, Cat:3211, ScienCell). All cell lines were cultured in a 10 cm culture dish (Corning) in a humidified incubator at 37 °C with 5% CO₂. The cells were harvested using 0.25% Trypsin/EDTA (Corning) when they reached 85−90% confluence.

Luo S, & Chen M. (2023). Systematic Investigation of the Effect of Lactobacillus acidophilus TW01 on Potential Prevention of Particulate Matter (PM)2.5-Induced Damage Using a Novel In Vitro Platform. Foods, 12(17), 3278.

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DENV Virus Propagation and Quantification

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DENV stock was obtained in vitro by infecting the C6/36 cell line (from Aedes albopictus larvae) with brain extracts of infected neonate mice. C6/36 cells were grown in minimum essential medium eagle (MEM) supplemented with 10% Fetal Bovine Serum (Gibco, NY, USA), Amphotericin B, Penicillin, Streptomycin, Pyruvate, Vitamins and l-glutamine, at 34 °C in 75-cm² culture flask (Corning, NY, USA). Infection was performed when cells reached 95% of confluency. After 48 h of infection, culture supernatant containing DENV was collected and concentrated with Amicon Centrifugal Filter Units (Merk Millipore, MA, USA). Infectious virion quantification was performed using a plaque-forming assay in Monkey African Green kidney cell line (Vero) and reported as Plaque-Forming Units (PFU)/mL.

Marcial-Juárez E., García-Cordero J., Maqueda-Alfaro R.A., Saucedo-López R.E., Sánchez-Torres L.E., Cedillo-Barrón L, & Flores-Romo L. (2020). Cutaneous Dengue Virus Inoculation Triggers Strong B Cell Reactions but Contrastingly Poor T Cell Responses. Virologica Sinica, 35(5), 575-587.

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Culturing J774A.1 Macrophages

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J774A.1 macrophages were grown at 37ºC (5% CO₂) in DMEM (Corning CellGro) containing 10% fetal bovine serum (Thermo Scientific), 1 mM pyruvate, 2 mM L-glutamine, and 1 mM penicillin/streptomycin (Corning CellGro). Cells were grown until confluent, scraped, and plated in medium lacking antibiotics at either 1 × 10⁵ cells/mL. Cells were allowed to adhere overnight before addition of experimental treatments.

Coulson G.B., Johnson B.K., Zheng H., Colvin C.J., Fillinger R.J., Haiderer E.R., Hammer N.D, & Abramovitch R.B. (2017). Targeting Mycobacterium tuberculosis sensitivity to thiol stress at acidic pH kills the bacterium and potentiates antibiotics. Cell chemical biology, 24(8), 993-1004.e4.

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Maintenance and Starvation of MEF and U2OS Cells

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Cells were maintained at 37°C in a humidified incubator at 5% CO₂. MEFs were seeded ~48h prior to the beginning of stress/starvation treatments and were maintained in DMEM (1.0g/l glucose, Cellgro, Manassas, VA) supplemented with 10% (v/v) FBS (Gibco, Grand Island, NY) and penicillin/streptomycin (Sigma, St. Louis, MO). MEFs were seeded at 5×10⁵ in 100mm plates.
U2OS cells were plated for ~72h prior to being fed the starvation media. U2OS cells were maintained in DMEM (4.5g/l glucose; Cellgro) supplemented with 10% (v/v) FBS and penicillin/streptomycin. U2OS were seeded at 1×10⁶ in 150mm plates. Starvation media utilized glucose free DMEM (0g/l glucose; Gibco) supplemented with 10% (v/v) dialyzed FBS (Gibco) and penicillin/streptomycin. The starvation media was added when the cells were between 85%-95% confluence. All media used either contained pyruvate or was supplemented with pyruvate (Cellgro) at a final concentration of 1mM. Starvation experiments utilized a variety of different forms of FBS including: Complete FBS (Gibco), delipidated FBS (Cocalito Biologicals; Reamstown, PA), and charcoal stripped FBS (Gibco).

Reeves R.A., Lee A., Henry R, & Zachara N.E. (2014). Characterization of the Specificity of O-GlcNAc Reactive Antibodies Under Conditions of Starvation and Stress. Analytical biochemistry, 457, 8-18.

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