Glucose

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Glucose is a monosaccharide and the primary source of energy for the body's cells. It is a key component in various laboratory and medical applications, serving as a critical analyte in the measurement of metabolic processes and disorders.

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Lab products found in correlation

Glucose, by Merck Group (3 mentions) Hepes, by Merck Group (2 mentions) Adenine hemisulfate, by MP Biomedicals (2 mentions) Complete supplement mixture without tryptophan, by MP Biomedicals (2 mentions) Eby100, by Thermo Fisher Scientific (2 mentions) C thermocellum dsm1237, by American Type Culture Collection (2 mentions) C cellulovorans, by American Type Culture Collection (2 mentions) C cellulolyticum, by American Type Culture Collection (2 mentions) R flavefaciens, by American Type Culture Collection (2 mentions) Mach1, by Thermo Fisher Scientific (2 mentions) Ammonium sulfate, by BD (2 mentions) Recombinant eby100, by BD (2 mentions) Acetosyringone, by Merck Group (2 mentions) Nah2po4, by Merck Group (2 mentions) Peptone, by Merck Group (1 mentions) Hepes, by MP Biomedicals (1 mentions) Peptone, by Lab M (1 mentions) 25 cm3 flasks, by Corning (1 mentions) Hypoxanthine, by Merck Group (1 mentions) Gentamicin, by Thermo Fisher Scientific (1 mentions)

10 protocols using glucose

Fluconazole Susceptibility Determination

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The fluconazole susceptibilities of the strains were determined by a previously described broth microdilution method (63 (link)), with slight modifications. A 2-day-old colony from a YPD agar plate was suspended in 2 ml of an 0.9% NaCl solution, and 4 µl of the suspension was mixed with 2 ml 2× SD-CSM medium (13.4 g yeast nitrogen base without amino acids [YNB; MP Biomedicals, Illkirch, France], 40 g glucose, 1.58 g complete supplement medium [MP Biomedicals] per liter). A 2-fold dilution series of fluconazole (Sigma GmbH, Deisenhofen, Germany) was prepared in water, starting from an initial concentration of 512 µg/ml. One hundred microliters of each fluconazole solution was then mixed with 100 µl of the cell suspension in a 96-well microtiter plate, and the plates were incubated for 48 h at 37°C. The MIC of fluconazole was defined as the drug concentration that abolished or drastically reduced visible growth compared to a drug-free control.

Popp C., Ramírez-Zavala B., Schwanfelder S., Krüger I, & Morschhäuser J. (2019). Evolution of Fluconazole-Resistant Candida albicans Strains by Drug-Induced Mating Competence and Parasexual Recombination. mBio, 10(1), e02740-18.

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Yeast Culture and Recombinant Antifungal Protein Production

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The strains used in this study are listed in Table 1. Yeast cells were cultured in the following liquid media: YPD (yeast extract (10 g/L; LabM, UK), peptone (20 g/L; LabM, UK) and glucose (20 g/L; Sigma-Aldrich, USA), YNB (yeast nitrogen base without amino acids; MP Biomedicals, USA) (6.7 g/L), PDB/YPD (potato dextrose broth (19.2 g/L; BD, USA), yeast extract (2 g/L), peptone (4 g/L) and glucose (4 g/L)) adjusted to pH 7 with 50 mM HEPES (Sigma-Aldrich, USA), synthetic complete (SC) medium (CSM (complete amino acid supplement mixture; MP Biomedicals, USA) (0.77 g/L), YNB (6.7 g/L) and glucose (20 g/L)) adjusted to pH 7 with 50 mM HEPES or 1/5^th PDB/YNB; at 30°C and 37°C for S. cerevisiae and C. albicans, respectively. Yeast cells were grown on following solid media: YPD agar plates containing 2% or 0.05% glucose (YPD + 15 g/L agar (Invitrogen, USA)) for all FC50 and replicative lifespan experiments. HsAFP1 and HsAFP1[H32A][R52A] were produced recombinantly, as described previously [4 (link), 11 (link)].

Struyfs C., Cools T.L., Cremer K.D., Sampaio-Marques B., Ludovico P., Wasko B.M., Kaeberlein M., Cammue B.P, & Thevissen K. (2020). The antifungal plant defensin HsAFP1 induces autophagy, vacuolar dysfunction and cell cycle impairment in yeast. Biochimica et biophysica acta. Biomembranes, 1862(8), 183255.

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Malaria Parasite Culture and Proliferation Assay

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The strains 3D7, W2, and Dd2 of P. falciparum malaria parasites (BEI Resources, MR4/ATCC, Manassas, VA, USA) were cultured in human type O+ erythrocytes in complete medium (RPMI 1640 (Cellgro), 0.043 mg/mL gentamicin (Gibco), 0.014 mg/mL hypoxanthine (Acros), 38.5 mM HEPES (Sigma), 0.18% NaHCO₃ (Cellgro),0.20% Glucose (MP Biomedical), 0.003 mM NaOH (Sigma), 0.2% Albumax (Gibco), 5% human serum) as previously described.^{14 (link)} Briefly, cultures were maintained in 25 cm³ flasks (Corning) at a volume of 10 mL and were gassed for 30 s with an environment of 3% CO₂, 1% O₂, and 96% N₂, then incubated at 37 °C.
The antimalarial activity was determined with a SYBR Green based parasite proliferation assay, adapted from Smilkstein^{15 (link)} and Bennett.^{16 (link)} Using this technique, the increase of parasite DNA contained in human red blood cells was measured after 72 h of incubation in the presence of serial dilutions of compounds.^{17 (link)} Relative fluorescence values were measured with a Molecular Devices SpectraMAX Gemini EM fluorimeter (excitation 495 nm and emission 525 nm). Data were analyzed using Microsoft Excel and plotted with SigmaPlot 10 (Systat).

de Melo Burger M.C., Fernandes J.B., das Graças Fernandes da Silva M.F., Escalante A., Prudhomme J., Le Roch K.G., Izidoro M.A, & Vieira P.C. (2014). Structures and Bioactivities of Dihydrochalcones from Metrodorea stipularis. Journal of natural products, 77(11), 2418-2422.

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Yeast and Bacterial Strains for Protein Expression

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The yeast S. cerevisiae strain EBY100 (Invitrogen, Carlsbad, CA) was used for mSEA surface display. The E. coli strains (Invitrogen) Mach1 and BL21 (DE3) were used for recombinant DNA manipulation and protein expression, respectively. All recombinant yeast and E. coli strains are summarized in Supplementary Table 1. C. thermocellum DSM1237, C. cellulovorans, C. cellulolyticum and R. flavefaciens were purchased from ATCC (Manassas, VA) and cultured anaerobically following ATCC protocols. Recombinant EBY100 cells were cultured using SC-Trp medium: 1.67 g/L yeast nitrogen base without amino acids, 5 g/L ammonium sulfate (Difco Laboratories, Detroit, MI), 20 g/L glucose, 15 g/L adenine hemisulfate, and 0.64 g/L complete supplement mixture without tryptophan (MP Biomedicals, Solon, OH). Induction of aScaf display on yeast surface was performed in YPG media (1% yeast extract, 2% peptone, 2% galactose). E. coli was cultured in Luria-Bertani (LB) medium containing 50 μg/mL kanamycin. Unless otherwise indicated, all chemicals were purchased from Sigma-Aldrich (St. Louis, MO).

Smith M.R., Gao H., Prabhu P., Bugada L.F., Roth C., Mutukuri D., Yee C.M., Lee L., Ziff R.M., Lee J.K, & Wen F. (2019). Elucidating structure-performance relationships in whole-cell cooperative enzyme catalysis. Nature catalysis, 2(9), 809-819.

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Drosophila Microbiome Isolation and Characterization

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Drosophila melanogaster Canton S (Wolbachia-free) were reared at 25°C, 12 h:12 h light-dark cycle, on yeast-glucose diet: 100 g l⁻¹ Brewer's yeast (inactive; MP Biomedicals), 100 g l⁻¹ glucose (Sigma), 12 g l⁻¹ agar (Apex) and preservatives [0.04% phosphoric acid, 0.42% propionic acid (Sigma)]. Drosophila gut microbiota members were isolated on modified MRS agar from aseptically-dissected fly guts. All bacteria used in the study are listed in Table 1, and were maintained at 30°C. Modified MRS contains (all from Sigma unless noted): 1.25% vegetable peptone (Becton Dickinson), 0.75% yeast extract, 2% glucose, 0.5% sodium acetate, 0.2% dipotassium hydrogen phosphate, 0.2% triammonium citrate, 0.02% magnesium sulfate heptahydrate, 0.005% manganese sulfate tetrahydrate, 1.2% agar (Apex). Potato medium contains: 0.5% glucose, 1% yeast extract, 1% peptone, 0.8% potato extract (Fluka 07915), 1.2% agar (Apex). Bacteria we isolated from Drosophila guts are identified as ^F for “fly isolate” and those isolated from other sources as ^NF for “non-fly,” e.g., Acetobacter tropicalis^F vs. A. tropicalis^NF.

Newell P.D., Chaston J.M., Wang Y., Winans N.J., Sannino D.R., Wong A.C., Dobson A.J., Kagle J, & Douglas A.E. (2014). In vivo function and comparative genomic analyses of the Drosophila gut microbiota identify candidate symbiosis factors. Frontiers in Microbiology, 5, 576.

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Culturing Cells for AMH Treatment

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GN11, GT1-7 and COS-7 cells were grown in monolayers at 37°C under 5% CO₂, in DMEM (ThermoFisher, Invitrogen) containing 1 mM pyruvate, 2 mM L-glutamine (ThermoFisher, Invitrogen), 100 μg/ml streptomycin, 100 U/ml penicillin and 9 mg/ml glucose (MP Biomedicals, Santa Ana, CA), supplemented with 10% fetal bovine serum (complete medium). Cells were maintained below full confluence by trypsination and seeding onto 10 cm² dishes. Cells used for experiments were between their third and eighth passage. Cells were treated with recombinant human AMH (1737-MS; R&D systems) at the concentrations ranging from 10 ng/ml to 250 ng/ml.

Malone S.A., Papadakis G.E., Messina A., Mimouni N.E., Trova S., Imbernon M., Allet C., Cimino I., Acierno J., Cassatella D., Xu C., Quinton R., Szinnai G., Pigny P., Alonso-Cotchico L., Masgrau L., Maréchal J.D., Prevot V., Pitteloud N, & Giacobini P. (2019). Defective AMH signaling disrupts GnRH neuron development and function and contributes to hypogonadotropic hypogonadism. eLife, 8, e47198.

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Yeast and Bacterial Strains for Protein Expression

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Engineered Y. lipolytica Strains for Research

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All the strains used in this study were constructed in the Y. lipolytica PO1f background (ATCC MYA-2613), a leucine and uracil auxotroph devoid of any secreted protease activity [39 ]. The ku70-disrupted Y. lipolytica strain was constructed using the ku70 deletion cassettes with URA3 marker as described by Verbeke [31 (link)]. All the strains used in this study are listed in Table 1.
Media and growth conditions for Escherichia coli and Y. lipolytica have been previously described by Sambrook and Russell [41 ], and Barth and Gaillardin [42 (link)], respectively. While, the Yeast Extract Peptone Dextrose (YPD) medium was prepared with 20 g/L bacto peptone (Difco Laboratories), 10 g/L yeast extract (Difco), 20 g/L glucose (Sigma-Aldrich), the Yeast Nitrogen Base (YNB) medium was made with 6.7 g/L yeast nitrogen base (without amino acids) (Difco), 0.69 g/L complete amino acid supplement mixture (CSM)-uracil supplement (MP Biomedicals), and 20 g/L glucose. HU was obtained from Sigma-Aldrich.

Jang I.S., Yu B.J., Jang J.Y., Jegal J, & Lee J.Y. (2018). Improving the efficiency of homologous recombination by chemical and biological approaches in Yarrowia lipolytica. PLoS ONE, 13(3), e0194954.

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Enzymatic Characterization of MAX1 Variants

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Enzymatic characterization of MAX1 and the genetic variants was carried out as described in Zhang et al. (2014) except that Agrobacterium tumefaciens was resuspended in 50 mM MES (Duchefa, Haarlem, The Netherlands)-KOH buffer (pH 5.6) containing 2 mM NaH 2 PO 4 (Merck, Darmstadt, Germany), 100 µM acetosyringone (Sigma-Aldrich, St. Louis, MO, USA), and 0.5% (w/v) glucose (MP Biomedicals, France) to a final OD 600 of 0.5. Instead of OsD27, OsCCD7, and OsCCD8, we used AtD27, MAX3, and MAX4, which were co-infiltrated with AtMAX1 and genetic variants to study the conversion of CL to CLA. Infiltration was performed using 4-week-old N. benthamiana plants which were soil grown in pots in a plant house with artificial light to make a photoperiod of 16 h light at 25 °C and 8 h dark at 22 °C. For each gene combination, six individual plants were used as biological replicates.

Xu X., Jibran R., Wang Y., Dong L., Flokova K., Esfandiari A., McLachlan A.R.G., Heiser A., Sutherland-Smith A.J., Brummell D.A., Bouwmeester H.J., Dijkwel P.P, & Hunter D.A. (2021). Strigolactones regulate sepal senescence in Arabidopsis. Journal of experimental botany, 72(15).

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Transient Expression of MAX1 Variants

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To characterise the enzymatic activity of MAX1 and the genetic variants described herein we used transient expression in N. benthamiana essentially as described in Zhang et al. (2014) except that A. tumefaciens were resuspended in 50 mM MES (2-Morpholinoethanesulfonic acid hydrate) (Duchefa, Haarlem, The Netherlands) -KOH buffer (pH 5.6) containing 2mM NaH 2 PO 4 (Merck, Darmstadt, Germany), 100 µM acetosyringone (Sigma-Aldrich, St. Louis, MO, USA) and 0.5% glucose (MP Biomedicals, France) to a final OD 600 of 0.5. Instead of OsD27, OsCCD7 and OsCCD8 that were shown by Zhang et al. (2014) to result in the production of CL upon transient expression in N. benthamiana we used AtD27, MAX3 and MAX4. By co-infiltrating MAX1 and genetic variants with these genes we can study the conversion of CL to CLA. Infiltration was performed using 4week-old N. benthamiana plants. For each gene combination, six individual plants were used as biological replicates.

Xu X., Jibran R., Wang Y., Dong L., Floková K., Esfandiari A., McLachlan A., Heiser A., Sutherland‐Smith A.J., Bouwmeester H.J., Dijkwel P.P., & Hunter D.A. (2020). Strigolactone defective mutants of Arabidopsis exhibit delayed sepal senescence.

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