PCE, cDCE (both ≥99.5%), VC (≥99.5%), ethene (≥99.9%), Vitamin B12 (≥98%), sodium fumarate (≥98%), and N2O (≥99%) were purchased from Sigma-Aldrich (St Louis, MO, USA). All other chemicals used in this study were reagent grade or of higher purity.
Sodium fumarate
Manufactured by Merck Group
Sourced in United States
Sodium fumarate is a chemical compound used in laboratory equipment. It is a white crystalline solid that is soluble in water. Sodium fumarate is used as a reagent and a source of the fumarate ion in various chemical reactions and analyses.
Automatically generated - may contain errors
Lab products found in correlation
Sodium nitrate, by Merck Group (2 mentions)
Ethene, by Merck Group (1 mentions)
Vitamin b12, by Merck Group (1 mentions)
Trimethylamine n oxide tmao, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Sodium succinate, by Merck Group (1 mentions)
Gaspak sachets system, by BD (1 mentions)
Sodium azide, by Merck Group (1 mentions)
Sodium formate, by Merck Group (1 mentions)
Sodium cyanide, by Merck Group (1 mentions)
L malic acid, by Merck Group (1 mentions)
Paraquat dichloride hydrate, by Merck Group (1 mentions)
Benzyl viologen dichloride, by Merck Group (1 mentions)
Catalase from bovine liver, by Merck Group (1 mentions)
Brain heart infusion agar, by Carl Roth (1 mentions)
Hemin, by Merck Group (1 mentions)
Metronidazole, by Merck Group (1 mentions)
Xanthine, by Merck Group (1 mentions)
Anaerocult a, by Merck Group (1 mentions)
7 protocols using sodium fumarate
1
Synthesis and Purification of Key Reagents
2
Bacterial Growth Curve Optimization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacterial strains were grown on LB streptomycin (100μg/ml) agar media and after 16-18h used to inoculate 4ml LB media. After 16h, bacterial strains were concentrated to a 1.0 OD600. 700μl LB media was inoculated 1:1000 (0.7μl) with the 1.0 OD600 resuspensions, vortexed, and aliquoted in triplicate 200μl volumes in a 96-well plate. Optical density was recorded every hour for the duration of the growth curve. Deoxygenated LB was used for anaerobic growth curves and benchtop LB used for aerobic growth curves. Concentrations of alternative electron acceptors supplemented to LB media were as follows: 50mM sodium fumarate (Sigma), 50mM trimethylamine-N-oxide (TMAO) (Sigma), 50mM sodium nitrate (Sigma), and 50mM dimethyl sulfoxide (DMSO) (Sigma). For strains grown in 50mM LB Nitrate media, after 3h, 5μM sodium hydroxide (Fisher Chemical) final concentration was added to alkalinize the growth media to support continued nitrate respiration.
3
Culturing Campylobacter jejuni under aerobic and anaerobic conditions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C. jejuni 81–176 (wildtype)24 (link) was cultured on Mueller-Hinton (MH) agar at 42 °C under microaerobic conditions (85% N2, 10% CO2, 5% O2) or under <1% O2 (hereafter referred to as anaerobic conditions) which was generated using the BD GasPak Sachets system (BD Diagnostics, NJ, USA)11 (link). The Campylobacter selective supplement (SR155E, Oxoid, KS, USA), sodium formate, sodium succinate, sodium fumarate, sodium cyanide, sodium azide and sodium nitrate (Sigma, MO, USA) were added to the MH medium when necessary.
4
Quantifying Metabolic Profiles in VAT Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The metabolite contents of the collected VAT culture media were determined by 1H-NMR, as previously described [20 (link)]. As an internal standard, 1 mM of sodium fumarate (Sigma-Aldrich, St. Louis, MO, USA) was used (singlet, 6.50 ppm).
Spectra analysis enabled us to identify and quantify the following metabolites (multiplicity, chemical shift): H1-α-glucose (doublet, 5.22 ppm), pyroglutamate (doublet of doublets, 4.16 ppm), pyruvate (singlet, 2.38 ppm), acetate (singlet, 1.90 ppm), alanine (doublet, 1.44 ppm), lactate (doublet, 1.33 ppm), isoleucine (doublet, 0.99 ppm) and valine (doublet, 0.97 ppm). The relative areas of 1H-NMR resonances were quantified using peak area integration with NUTS-Pro (Acorn NMR, Livermore, CA, USA). The results are expressed as nanomoles of metabolite consumed/produced per milligram of WVAT.
Spectra analysis enabled us to identify and quantify the following metabolites (multiplicity, chemical shift): H1-α-glucose (doublet, 5.22 ppm), pyroglutamate (doublet of doublets, 4.16 ppm), pyruvate (singlet, 2.38 ppm), acetate (singlet, 1.90 ppm), alanine (doublet, 1.44 ppm), lactate (doublet, 1.33 ppm), isoleucine (doublet, 0.99 ppm) and valine (doublet, 0.97 ppm). The relative areas of 1H-NMR resonances were quantified using peak area integration with NUTS-Pro (Acorn NMR, Livermore, CA, USA). The results are expressed as nanomoles of metabolite consumed/produced per milligram of WVAT.
5
Bacterial Biofilm Quantification Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacterial cultures were normalized to an OD600 of 1.0 and inoculated at 1:100 into TSB supplemented with 0.5% glucose in a 96-well plate. Sodium fumarate (Sigma-Aldrich) and L-malic acid (Sigma-Aldrich) were used at final concentrations of 62.5 mM, 125 mM, 250 mM, and 500 mM in TSB containing 0.5% glucose. Biofilm cultures were grown statically at 37°C. After 24 hours, OD600 was measured and plates were washed twice with water, dried, and fixed with methanol. They were then stained with 1% crystal violet (wt/vol) and again washed twice with water and dried. After solubilization of the well contents with 33% acetic acid (vol/vol), OD540 values were determined.
6
Anaerobic Microbial Growth Enablers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wilkins-Chalgren anaerobe agar (WC) was purchased from Oxoid (Basingstoke, England), and Brain Heart Infusion Broth (BHI), Brain Heart Infusion Agar, and vitamin K1 were purchased from Carl Roth (Karlsruhe, Germany). Hemin, metronidazole, NADH, NADPH, cytochrome c, benzyl viologen dichloride, catalase from bovine liver, paraquat dichloride hydrate, Tris/HCl, Triton X-100, xanthine, pyruvic acid, oxaloacetate, sodium fumarate, β-mercaptoethanol, and Coenzyme A were all purchased from Sigma-Aldrich (St. Luis, USA). Potassium dihydrogen phosphate (KH2PO4), hydrogen peroxide, sodium dithionite, xanthine oxidase, ethylenediaminetetraacetic acid (EDTA), sodium chloride, and Anaerocult A were purchased from Merck (Darmstadt, Germany). Etests were purchased from bioMérieux (Marcy-l'Étoile, France).
7
Anaerobic growth and fumarate impact
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anaerobiosis was achieved using a Coy Anaerobic Chamber. For LB growth curves, a 1:1000 dilution of a 1.0 OD600 culture was used to inoculate prepared media which was grown statically at 37ºC. WT, ΔaceE, and ΔaceF growth curves were performed in 50mL of LB media in a 125mL flask, whereas later WT and ΔpflA LB growth curves were performed in 2mL of LB media in a 15mL round bottom tube. For M9 + 0.5% PSIM, 2mL media was added to a 15mL round bottom tube and inoculated 1:250 with 1.0 OD600 culture and grown statically at 37ºC. At each timepoint, flask and tubes were swirled or vortexed and 100µl was removed for dilution series plating. For growth curves including 50mM fumarate, sodium fumarate (Sigma) reagent was used.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!