Cells were grown in yeast complete media lacking both uracil and lysine (YC/Ura−Lys−) plus 2% raffinose (Sigma), diluted into YC/Ura− plus 2% raffinose and cultured to logarithmic phase. Galactose (Sigma) was added to a final concentration of 3%, and cells were cultured for an additional 24 hrs. Samples were processed for plating or dotting assays. Both liquid and solid cultures were grown at 30°C.
Raffinose
Manufactured by Merck Group
Sourced in United States, Germany, China, Switzerland, United Kingdom
Raffinose is a trisaccharide sugar that consists of galactose, fructose, and glucose. It is commonly used in laboratory settings as a carbohydrate source and for the cultivation of microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Glucose, by Merck Group (31 mentions)
Sucrose, by Merck Group (26 mentions)
Fructose, by Merck Group (22 mentions)
Galactose, by Merck Group (22 mentions)
Stachyose, by Merck Group (20 mentions)
Melibiose, by Merck Group (12 mentions)
Lactose, by Merck Group (11 mentions)
Maltose, by Merck Group (10 mentions)
D galactose, by Merck Group (7 mentions)
D glucose, by Merck Group (6 mentions)
Myo inositol, by Merck Group (6 mentions)
Arabinose, by Merck Group (6 mentions)
Gallic acid, by Merck Group (5 mentions)
Rhamnose, by Merck Group (5 mentions)
Trehalose, by Merck Group (5 mentions)
Verbascose, by Merck Group (5 mentions)
Cellobiose, by Merck Group (4 mentions)
Caffeic acid, by Merck Group (4 mentions)
Mannose, by Merck Group (4 mentions)
D fructose, by Merck Group (4 mentions)
88 protocols using raffinose
1
Yeast Growth Induction Assay
2
Quantification of Isoflavones and Carbohydrates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Standard solutions of minerals were purchased from Quimlab Química & Metrologia® (São Paulo, Brazil). Commercial standards of stachyose and sucrose were purchased from Sigma (St. Louis, MO, USA), raffinose from Merck (Darmstadt, Germany) and daidzin, glycitin, genistin, daidzein, glycitein and genistein from Apin Chemicals Limited® (Abingdon, UK). A commercial food grade fungal cellulase enzyme (CelluMax C) was donated from Prozyn (São Paulo, Brazil). All solvents were HPLC grade from Tedia (Fairfield, OH, USA) and HPLC grade water (Milli-Q system, Millipore, Bedford, MA, USA) was used throughout the experiments.
3
Quantitative Analysis of Plant Sugars
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sugars were extracted as previously described (Muir et al. 2009 (link)) with minor changes. Briefly, 100 mg of powder was mixed with 8 mL distilled water in a 10-mL volumetric flask by sonication for 15 min at 80 °C in a Branson 3510 device (Marshal Scientific, Hampton, NH, USA). After cooling to room temperature (25 °C), the volume was adjusted to 10 mL with distilled water and the sample was centrifuged (3000 ×g, 10 min, room temperature). The supernatant was passed through a 0.22-µm sterile Millex filter (Merck, Overijse, Belgium) and analyzed by size exclusion chromatography (SEC) using an Acquity H-Class ultra-high performance liquid chromatography (UPLC) system (Waters, Milford, MA, USA) with a refractive index detector and external column oven. We used two TSK gel G2500PWXL columns (Tosoh, Tokyo, Japan) at 80 °C and a guard column (Bio-Rad, Temse, Belgium) under isocratic conditions, with distilled water as the mobile phase at a flow rate of 0.5 mL/min. The injected volume was 10 µL, and each run lasted 40 min. Sugars were identified by retention time and quantified against calibration curves of sucrose, fructose, glucose, raffinose and stachyose standards (Merck).
4
Antioxidant Capacity Evaluation Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fast Blue BB (FBBB) [4-benzoylamino-2,5-dimethoxybenzenediazonium chloride hemi-(zinc chloride) (Rebello et al., 2016 (link)), bile extract porcine, pancreatin from porcine pancreas, pepsin from porcine gastric mucosa, α-amylase from human saliva, 2,2′-azinobis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2′-diazobis-(2-aminodinopropane)-dihydrochloride (AAPH), fluorescein, 2,2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich, Co. (St. Louis, MO, United States). Standards of chlorogenic acid, 2-coumaric acid, 2,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, vanillic acid, and vanillin were provided by Extrasynthese (Lyon, Genay Cedex, France). Standards of 6-hydroxy-2,5,7, 8-tetramethyl-2-carboxylic acid (Trolox), gallic acid, caffeic acid, ferulic acid, vitexin, D-(+)-glucose, D-(+)-galactose, D-(+)-xylose, D-(–)-arabinose, and stachyose were acquired from Sigma-Aldrich, Co. (St. Louis, MO, United States). Standards of 33-α-L-plus 23-α-L-arabinofuranosyl-xylotetraose (XA3XX/XA2XX), 23,33-di-α-L-arabinofuranosyl-xylotriose (A2, 3XX), and xylotriose were obtained from Megazyme (Wicklow, Ireland). Raffinose and cellobiose standards were supplied by Merck (Darmstadt, Germany).
5
Analytical Standards Characterization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LC-MS grade methanol (Supelco LiChroSolv hypergrade) and fuming hydrochloric acid (37% HCl) were obtained from Merck Life Science AB, UK. Acetonitrile (ACN) was obtained from Sigma-Aldrich (Supelco LiChroSolv hypergrade for LC-MS) and formic acid (FA) was obtained from VWR (AmalaR Normapur, VWR Sweden). Suwannee River natural organic matter (SRNOM) was obtained from the International Humic Substances Society (IHSS, Saint-Paul, USA; batch number 2R101N). Hippuric acid, fusidic acid, cyclohexyl succinic acid, capsaicin, adenosine-5-monophosphate, raffinose, carbenoxolone disodium and glycyrrhizic acid were obtained from Merck Life Science AB, Sweden.
6
Enzymatic Assay for α-Galactosidase Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
p-Nitrophenyl α-galactopyranoside (α-p-NPGal) was purchased from Sigma Chemicals Company. Agar was provided by Fluka, Spain. 2, 4, 6-Tri (2-pyridyl)-s-triazine (TPTZ), 1,1′-diphenyl-2-picrylhydrazyl (DPPH), butylated hydroxyanisole (BHA), tert-butylated hydroxyl quinone (TBHQ), galactose, glucose, fructose, sucrose, raffinose, and stachyose were purchased from Merck Chemical Company. All the other chemicals used were of analytical grade.
7
Mass Spectrometry Calibration and Sample Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Agilent ESI-L Low Concentration
Tune Mix containing fluoroalkyl phosphazene calibrant ions was used
as received from the vendor. Peptides (SDGRG and GRGDS) and trisaccharides
(melezitose, raffinose, maltotriose, and isomaltotriose) were purchased
from Millipore-Sigma (St. Louis, MO). Gangliosides (GD1a d18:1–18:0 and GD1b d18:1–18:0) were purchased
from Matreya, LLC (State College, PA). Triglycerides (1,2,3-tri-γ-linolenoyl-rac-glycerol and 1,2,3-tri-α-linolenoyl-rac-glycerol) were purchased from Cayman Chemical (Ann Arbor, MI). A
complete list of chemical standards and their sources can be found
in theSupporting Information (Table S1 ). All chemical standards except the
Triglycerides were obtained as dry powders and prepared individually
to final working solution concentrations of ∼10 μg/mL
in 1:1 methanol/water for IM-MS analysis. The Triglycerides were obtained
as 10 mg/mL solutions in ethanol and further prepared at 20 μM
in 1:1 methanol/isopropanol with 13 μM ammonium acetate. All
solvents were obtained from Fisher Scientific (Optima grade, Hampton,
NH).
Tune Mix containing fluoroalkyl phosphazene calibrant ions was used
as received from the vendor. Peptides (SDGRG and GRGDS) and trisaccharides
(melezitose, raffinose, maltotriose, and isomaltotriose) were purchased
from Millipore-Sigma (St. Louis, MO). Gangliosides (GD1a d18:1–18:0 and GD1b d18:1–18:0) were purchased
from Matreya, LLC (State College, PA). Triglycerides (1,2,3-tri-γ-linolenoyl-rac-glycerol and 1,2,3-tri-α-linolenoyl-rac-glycerol) were purchased from Cayman Chemical (Ann Arbor, MI). A
complete list of chemical standards and their sources can be found
in the
Triglycerides were obtained as dry powders and prepared individually
to final working solution concentrations of ∼10 μg/mL
in 1:1 methanol/water for IM-MS analysis. The Triglycerides were obtained
as 10 mg/mL solutions in ethanol and further prepared at 20 μM
in 1:1 methanol/isopropanol with 13 μM ammonium acetate. All
solvents were obtained from Fisher Scientific (Optima grade, Hampton,
NH).
8
Yeast Strain Culturing Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Yeast strains used in this study are MATa cells with A364A background, and their genotypes are listed in Supplementary file 1 . Synthetic complete with low biotin (SC-Dex) was prepared by dissolving 1.4 g/l YNB-biotin (Sunrise Science Products, see Key resources table), 1.6 g/l BSM powder (Sunrise Science Products), 5 g/l ammonium sulfate (Fisher Scientific), 20 g/l dextrose (Fisher Scientific), and 0.6 nM D-Biotin (Invitrogen) in distilled water and filter sterilized. Synthetic complete medium with raffinose (SC-Raff) for GAL induction was prepared similarly with SD-Dex, but 20 g/l dextrose was replaced with 20 g/l raffinose (Millipore-Sigma).
9
Glucose and Galactose Assay of Yeast Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Strains were obtained as described in Lee et al., 2017 (link); Wang et al., 2015 (link). An initial set of 36 strains were assayed in a glucose gradient (1–0.0039%) with a constant background of 0.25% galactose. Strains DBVPG6765, CLIB324, L-1528, M22, W303, YIIC17-E5 were excluded from downstream analysis due to poor growth in our media conditions. Strain 378604X was also excluded due to a high basal expression phenotype that was an outlier in our collection. The genetic basis of this behavior is likely an interesting topic for follow-up studies. All experiments were performed in synthetic minimal medium (‘S’), which contains 1.7 g/L yeast nitrogen base (YNB) (BD, Franklin Lakes, NJ) and 5 g/L ammonium sulfate (EMD). In addition, D-glucose (EMD, Darmstadt, Germany), D-galactose (MilliporeSigma, St. Louis, MO), mannose (MilliporeSigma), glycerol (EMD), acetate (MilliporeSigma), and/or raffinose (MilliporeSigma) were added as a carbon source. Cultures were grown in a humidified incubator (Infors Multitron, Bottmingen, CH) at 30°C with rotary shaking 999 rpm (500 µL cultures in 1 mL 96-well plates).
10
Sperm Cryopreservation in Raffinose-Skim Milk Medium
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sperm cryopreservation was performed using a cryoprotective medium (CPM) [1 (link)]. The CPM consisted of 18% w/v raffinose (Sigma Aldrich, cat #R7630), 3% w/v skim milk (BD Diagnostics, cat # 232100), and 477 μM monothioglycerol (MTG, Sigma Aldrich, cat #M6145). Spermatozoa collected from the caudae epididymides and vasa deferentia were allowed to disperse from the tissues for 10 min at 37 °C. After loading into 0.25 mL French straws (IMV Technologies, L’Aigle, France), the straws were sealed, exposed to liquid nitrogen vapors for 10 min, and then stored in LN2.
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!