The largest database of trusted experimental protocols

6 protocols using d glucose 6 phosphate disodium salt hydrate

1

In vitro metabolism of reactive oxygen species

Check if the same lab product or an alternative is used in the 5 most similar protocols
The chemicals and reagents used in our experiment were obtained from the following sources. PIO (purity = 97%) standard was obtained from Toronto Research Chemicals (North York, Ontario, Canada). ROS labeled with deuterium (on the benzene ring; ROS-D4, purity = 96%) standard was obtained from BDG Synthesis Limited (Wellington, New Zealand). Human liver S9 fractions (20 mg/mL protein base) were obtained from Thermo Fisher Scientific (Runcorn, United Kingdom). ROS (purity ≥98%) standard, glucose-6-phosphate dehydrogenase (activity: 225 units/mg), MgCl2 (purity ≥98%), sodium phosphate monobasic monohydrate (purity ≥98%), sulfatase (activity: 11 units/mL), D-glucose 6-phosphate disodium salt hydrate (purity ≥98%), β-glucuronidase (activity >85,000 units/mL), β-nicotinamide adenine dinucleotide phosphate sodium salt hydrate (NADP, purity ≥98%), acetic acid (purity ≥99%), and sodium phosphate dibasic (purity ≥99%) were obtained from Sigma-Aldrich (St. Louis, MO, United States).
+ Open protocol
+ Expand
2

Biochemical Assay Reagents and Materials

Check if the same lab product or an alternative is used in the 5 most similar protocols
Purified bovine serum albumin (BSA), T6P dipotassium salt, uridine 5′-diphosphoglucose disodium salt from Saccharomyces cerevisiae (UDPG), d-glucose-6-phosphate disodium salt hydrate (G6P), and alkaline phosphatase from bovine intestinal mucosa (lyophilized powder, 10–30 defined enzyme activity (DEA) units/mL solid), penicillin G sodium salt, nystatin, IVM, 4-(2-hydroxyethyl)-piperazine-1-ethanesulfonic acid (HEPES), agarose, and ethidium bromide were purchased from Sigma-Aldrich (Germany and USA). Total RNA Kit, TranScriba Kit, and SYBR Green B PCR-MIX Taq were obtained from A&A Biotechnology (Poland). The components of 0.1 M acetic acid-ammonia buffer and 0.1 M phosphoric buffer (resp., pH 4.2 and pH 7.0) (acetic acid, ammonia, NaH2PO4, and Na2HPO4  × 7H2O2) and of Ascaris Ringer's Solution (ARS) medium (KCl, CaCl2  × 2H2O, MgCl2  × 6H2O, NaCl, and sodium acetate) were of high purity grade and were purchased from Chempur (Poland) and P.P.H. Stanlab (Poland).
The water used for the analysis was deionized with the use of a Direct-Q Ultrapure UV3 Water System (EMD Millipore, USA). The water, media, sodium saline, and surgical instruments were sterilized using a Classic Standard Prestige Medical autoclave (Ma-Je-R, Poland).
+ Open protocol
+ Expand
3

Capillary Electrophoresis Analysis of Phosphates

Check if the same lab product or an alternative is used in the 5 most similar protocols
α-D-Glucose 1-phosphate disodium salt hydrate (≥97%), DGlucose 6-phosphate disodium salt hydrate (≥98%), DFructose 6-phosphate disodium salt hydrate (≥98%), acetonitrile (HPLC graded), ammonium hydroxide (HPLC graded) were purchased from Sigma-Aldrich (St. Louis, MO). Dulbecco’s Modified Eagle Medium (DMEM) media with no glucose, glutamine or phenol red added was used without further purification. Nanopure water (18.2 MΩ cm) was obtained from a Barnstead Nanopure filtration system. Bare fused silica capillary with 41.5 μm i.d. and 104 μm o.d. was purchased from Polymicro Technologies (Phoenix, AZ).
+ Open protocol
+ Expand
4

Enzymatic Activities in BM-MSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
5 × 105 cells from each group of different age were used for the assessment of enzyme activities. The cells were homogenized in 200 μL of 250 mM sucrose(Sigma-Aldrich, St.Louis, MO), 50 mM HEPES (Sigma-Aldrich), 0,5 mM EDTA (Sigma Aldrich) and one tablet protease inhibitor cocktail (Roche, Mannheim, Germany ). Enzymes activities were determined using a SUNRISE spectrophotometer (TECAN, Mannedorf, Switzerland). Reaction rates of enzymes were determined by the increase or decrease in absorbance of NAD(P)H (Sigma-Aldrich, St.Louis, MO) at 340 nm at 37 °C. Lactate dehydrogenise (EC 1.1.1.27) was determined in BM-MSCs using 50 mM Trizma base (pH 7,4), 0,15 mM NADH and 5 mM sodium pyruvate (omitted for control) (all Sigma Aldrich, St.Louis, MO). Glucose-6-phosphate 1-deshydrogenase (EC 1.1.1.49) and 6-phosphogluconate dehydrogenase, decarboxylating (EC 1.1.1.343) was determined in BM-MSCs using 78 mM Trizma base, 5 mM MgCl2(pH 7,4), 0,1 mM NADP, 0,5 mM D-Glucose 6-phosphate disodium salt hydrate and 6-Phosphogluconic acid trisodium salt (omitted for control) (all Sigma-Aldrich, St.Louis, MO).
+ Open protocol
+ Expand
5

Assay for Glucose-6-Phosphate Dehydrogenase Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
Glucose-6-phosphate dehydrogenase (activity: 225 units/mg), MgCl2 (purity: ≥98%), d-glucose 6-phosphate disodium salt hydrate (purity: ≥98%), sulfatase (activity: 11 units/mL), β-glucuronidase (activity: >85000 units/mL), sodium phosphate monobasic monohydrate (purity: ≥98%), β-nicotinamide adenine dinucleotide phosphate sodium salt hydrate (NADP, purity: ≥98%), sodium phosphate dibasic (purity: ≥99%), and acetic acid (purity: ≥99%) were obtained from Sigma–Aldrich (St. Louis, MO, USA). PIO (purity = 97%), deuterium-labeled rosiglitazone (purity = 98%), and deuterium-labeled (on the benzene ring) PIO (D4-PIO, purity = 97%) were obtained from Toronto Research Chemicals (Toronto, Canada). Human liver S9 fractions (20 mg/mL protein base) were obtained from Thermo Fisher Scientific (Runcorn, UK).
+ Open protocol
+ Expand
6

Preparation and NMR Analysis of Carbohydrate Metabolites

Check if the same lab product or an alternative is used in the 5 most similar protocols
The reference compounds D-glucose (Glc), α-D-galactose 1phosphate dipotassium salt (Gal-1P), α-D-glucose 1-phosphate disodium salt hydrate (Glc-1P), D-glucose 6-phosphate disodium salt hydrate (Glc-6P), D-ribose 5-phosphate disodium salt hydrate (Rib-5P), uridine 5′-diphosphogalactose disodium salt (UDP-Gal), uridine 5′-diphosphoglucose disodium salt hydrate (UDP-Glc), uridine 5′-diphospho-N-acetylgalactosamine disodium salt (UDP-GalNAc), uridine 5′-diphospho-Nacetylglucosamine sodium salt (UDP-GlcNAc), uridine 5′diphosphoglucuronic acid trisodium salt (UDP-GluA), and creatine were all obtained from Sigma-Aldrich ® , Switzerland.
Twenty-millimolar reference solutions were prepared in D 2 O and D 2 O-based phosphate-buffered saline (PBS) containing 10 mM creatine as internal reference for chemical shift calibration.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!