Sugar nucleotide extraction and analysis were performed as described (27 (link)). Briefly, cells were pelleted by centrifugation, washed in ice-cold PBS, and lysed in 70% ethanol in the presence of 20 pmol of the GDP-glucose as an internal standard (Sigma). The lysate was centrifuged to remove insoluble material, and the supernatant was extracted with butan-1-ol to remove lipids. Sugar nucleotides were extracted from the resulting aqueous phase using EnviCarb graphitized carbon columns (Supelco) as described previously (67 (link)). The eluted sugar nucleotides were analyzed by multiple-reaction monitoring LC-MS/MS (27 (link)).
Gdp glucose
Manufactured by Merck Group
GDP-glucose is a laboratory reagent used in biochemical and molecular biology research. It serves as a substrate for enzymes involved in the synthesis and interconversion of nucleotide-sugars, which are essential components of various biomolecules. The core function of GDP-glucose is to provide a source of glucose for these enzymatic reactions, facilitating the study of carbohydrate metabolism and signaling pathways in biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Envicarb, by Merck Group (1 mentions)
Sodium chloride (nacl), by Avantor (1 mentions)
P 10 desalting column, by GE Healthcare (1 mentions)
Udp galactose, by Merck Group (1 mentions)
Triton x 100, by Thermo Fisher Scientific (1 mentions)
Udp glucose, by Merck Group (1 mentions)
4 2 hydroxyethyl 1 piperazineethanesulfonic acid hepes, by Avantor (1 mentions)
Udp n acetylglucosamine, by Merck Group (1 mentions)
Udp glucuronic acid, by Merck Group (1 mentions)
Adp glucose, by Merck Group (1 mentions)
Isopropyl 1 thio β d galactopyranoside iptg, by Thermo Fisher Scientific (1 mentions)
Yeast extract, by BD (1 mentions)
Tryptone, by BD (1 mentions)
2 protocols using gdp glucose
1
Sugar Nucleotide Extraction and Analysis
2
Glycosyltransferase Activity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Reagent grade chloroform, methanol, and silica
gel 60 thin-layer chromatography (TLC) plates (layer thickness of
150–200
μm) were obtained from EMD Chemicals Inc. (Gibbstown,
NJ). [32P]Pi, UDP-[U-14C]glucose,
UDP-[U-14C]galactose, and GDP-[U-14C]mannose
were from
PerkinElmer Life and Analytical
Sciences, Inc. (Waltham, MA). Yeast extract, agar, and tryptone were
from BD Biosciences. Sodium chloride and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic
acid (HEPES) were from VWR International (West Chester, PA). Triton
X-100 was from Thermo Fisher Scientific (Waltham, MA). Isopropyl 1-thio-β-d -galactopyranoside (IPTG) was from Invitrogen Corp. (Carlsbad,
CA). UDP-glucose, UDP-galactose, UDP-glucuronic acid, UDP-galacuronic
acid, ADP-glucose, GDP-glucose, UDP-N-acetylglucosamine,
and ADP-mannose were purchased from Sigma-Aldrich.
The P-10 desalting column was from GE. All other chemicals were reagent
grade and were purchased from either Sigma-Aldrich (St. Louis, MO)
or Mallinckrodt Baker, Inc. (Phillipsburg, NJ).
gel 60 thin-layer chromatography (TLC) plates (layer thickness of
150–200
μm) were obtained from EMD Chemicals Inc. (Gibbstown,
NJ). [32P]Pi, UDP-[U-14C]glucose,
UDP-[U-14C]galactose, and GDP-[U-14C]mannose
were from
PerkinElmer Life and Analytical
Sciences, Inc. (Waltham, MA). Yeast extract, agar, and tryptone were
from BD Biosciences. Sodium chloride and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic
acid (HEPES) were from VWR International (West Chester, PA). Triton
X-100 was from Thermo Fisher Scientific (Waltham, MA). Isopropyl 1-thio-β-
CA). UDP-glucose, UDP-galactose, UDP-glucuronic acid, UDP-galacuronic
acid, ADP-glucose, GDP-glucose, UDP-N-acetylglucosamine,
and ADP-mannose were purchased from Sigma-Aldrich.
The P-10 desalting column was from GE. All other chemicals were reagent
grade and were purchased from either Sigma-Aldrich (St. Louis, MO)
or Mallinckrodt Baker, Inc. (Phillipsburg, NJ).
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