TPs were extracted with boiling water in a solid-liquid ratio of 1:20 (w/v) for 6 h. After being filtered and centrifuged, three times the volume of ethanol was added to obtain a precipitate, and the precipitate was deproteinated using the Sevage method (12 (link)). The deproteinated part was extensively dialyzed against distilled water and then applied to a 60 mm × 240 m column of DEAE-cellulose (Sigma, USA) at a flow rate of 3.0 mL/min. The column was then eluted with distilled water, 50 mM NaCl, 150 mM NaCl, and 1M NaCl at 10 mM/min, successively, and the part eluted by 50 mM NaCl was collected and lyophilized: this was defined as the TPs for further study.
Deae cellulose
Manufactured by Merck Group
Sourced in United States, Germany
DEAE-cellulose is a type of ion-exchange chromatography media. It is composed of cellulose that has been derivatized with diethylaminoethyl (DEAE) groups. DEAE-cellulose is used for the purification and separation of biomolecules, such as proteins and nucleic acids, based on their ionic interactions with the DEAE functional groups.
Automatically generated - may contain errors
Lab products found in correlation
Sephadex g 100, by Merck Group (4 mentions)
Bovine serum albumin (bsa), by Merck Group (4 mentions)
Sephacryl s 300, by Merck Group (3 mentions)
Guaiacol, by Merck Group (3 mentions)
D xylose, by Merck Group (3 mentions)
Tween 20, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
D arabinose, by Merck Group (2 mentions)
D glucuronic acid, by Merck Group (2 mentions)
L rhamnose, by Merck Group (2 mentions)
D mannose, by Merck Group (2 mentions)
D galactose, by Merck Group (2 mentions)
D galacturonic acid, by Merck Group (2 mentions)
Superdex 200 column, by GE Healthcare (2 mentions)
Oleic acid, by Merck Group (2 mentions)
D fructose, by Merck Group (2 mentions)
D glucose, by Merck Group (2 mentions)
Palmitate, by Merck Group (2 mentions)
Sephadex g 50, by Merck Group (2 mentions)
38 protocols using deae cellulose
1
Extraction and Purification of Polysaccharides
2
Protein Purification via Chromatography
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Sephadex G-100, DEAE-cellulose and guaiacol were purchased from Sigma-Aldrich Co, St Louis, USA. Standard protein markers were purchased from Merck, Genei, India. Dyes were collected from local textile industry. All chemicals used were of the highest purity available and of the analytical grade.
3
Xanthine Oxidase Activity Assay
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Xanthine sodium salt, xanthine oxidase enzyme, cytochrome C from horse heart, nitroblue tetrazolium (NBT), dimethyl sulfoxide (DMSO), phenyl methosulfate (PMS), phenymethylsulfonyl fluoride (PMSF), 1,4 dithiothreitol (DTT), 1,10 phenanthroline, trypan blue dye, bovine serum albumin (BSA), blue dextran, crystal violet, Sephacryl S-300, DEAE cellulose, and kits of gel filtration molecular weight marker were product of Sigma Co. SDS molecular weight marker proteins were purchased from Pharmacia Co., DMEM, fetal bovine serum, HEPES buffer solution, RPMI-1640, gentamycin, and L-glutamine are purchased from Lonza, Belgium.
4
Extraction and Characterization of Medicinal Plant Pectins
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DEAE-cellulose, Sepharose CL-6B, Sephadex G25, and aminobenzamide (2AB) were purchased from Sigma-Aldrich. Endo-polygalacturonase (Endo-PG, EC 3.2.1.15 from Aspergillus niger) was purchased from Megazyme. All chemicals used were analytical grade and produced in China.
Homogalacturonan pectins were extracted from the following plants: Panax japonicus, Pseudostellaria heterophylla, Schisandra chinensis, Prunella vulgaris, Panax Notoginseng, Polygonum orientale, Anemarrhena asphodeloides, Kadsura longipedunculata, Isatis indigotica, Aconitum carmichaelii, Coptis chinensis, and Sophora flavescens.
Homogalacturonan pectins were extracted from the following plants: Panax japonicus, Pseudostellaria heterophylla, Schisandra chinensis, Prunella vulgaris, Panax Notoginseng, Polygonum orientale, Anemarrhena asphodeloides, Kadsura longipedunculata, Isatis indigotica, Aconitum carmichaelii, Coptis chinensis, and Sophora flavescens.
5
Wheat Bran-Based Enzyme Production
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Major chemicals such as 2,2' azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), Sephadex G-100, Diethylamino ethyl Cellulose (DEAE-cellulose), bovine serum albumin (BSA) and Folin-ciocalteau reagents were obtained from Sigma-Aldrich, Germany while malt extract agar (MAE) and potato dextrose agar (PDA) were obtained from JHD, China. Wheat bran was procured from a nearby market in Nsukka, Enugu State, Nigeria and ground to a particle size of 1 mm. The powdered wheat bran was stored in a dry container and was used as both an inducer and lignocellulosic support.
6
Purification of Rice K-1 Cell Enzymes
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The rice k-1 cell line established from Oryza sativa L. cv. Nipponbare was kindly gifted by H. Nishimura and the late Professor K. Kasamo (Research Institute for Bioresources, Okayama University). DEAE-cellulose was purchased from Sigma (St. Louis, MO, United States), and Butyl-TOYOPEARL and TSK-Gel G3000SWXL columns (0.78 × 30 cm) were purchased from Tosoh (Tokyo, Japan). A Shodex PH-814 column (0.8 × 7.5 cm) was purchased from Showa Denko (Tokyo, Japan). A Cosmosil 5C18-AR column (0.6 × 25 cm) was purchased from Nacalai Tesque (Kyoto, Japan). Ni-NTA His•Bind® resin and GST•BindTM resin were purchased from Novagen®. Authentic PA-sugar chains were prepared as described in our previous studies (Kimura et al., 1988 (link), 1997 (link), 2000 (link)).
7
Bioactive Compounds from Fireweed Leaves
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The fireweed leaves were collected in July 2017 from plants grown in Komi Republic (Latitude: 61′14″ S; Longitude: 50′24″ E), Russia. Plant material was botanically identified by Dr. Nina N. Shergina from Syktyvkar State University, Syktyvkar, Russia.
Reagents, including bovine serum albumin, catalase, cytochrome c, DEAE-cellulose, 3,5-dimethylphenol, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), the Folin and Ciocalteu’s phenol reagent, myo-inositol, 1,4-α-d -polygalacturonase, rhamnose (Rha), arabinose (Ara), galactose (Gal), mannose (Man), xylose (Xyl), glucose (Glc), galacturonic acid (GalA), sodium borohydride, sodium chloride, superoxide dismutase, xanthine and xanthine oxidase were purchased from Sigma-Aldrich (St. Louis, MO, USA). The pullulan standards were purchased from Fluka (Steinheim, Germany) and PSS Polymer Standards Service GmbH (Mainz, Germany). Gallic acid was purchased from MP Biomedicals (Solon, OH, USA).
Reagents, including bovine serum albumin, catalase, cytochrome c, DEAE-cellulose, 3,5-dimethylphenol, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), the Folin and Ciocalteu’s phenol reagent, myo-inositol, 1,4-α-
8
Purification of Trypsin Inhibitor from Gold Beans
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A water extract of gold beans Phaseolus vulgaris cv. “gold bean” from Mainland China (260 g) was made by homogenizing them in distilled water (6 mL/g). The homogenate was then centrifuged (14,000× g for 25 min at 4 °C). The supernatant was collected and loaded on a 5 × 20 cm column of DEAE-cellulose (Sigma, St. Louis, MI, USA) in 10 mM Tris-HCl buffer (pH 7.4). Following removal of unadsorbed proteins (fraction D1), the column was eluted sequentially with 0.2 M NaCl and 1 M NaCl in the Tris-HCl buffer. Fraction D2 eluted with 0.2 M NaCl was dialyzed and then chromatographed on a 5 × 15 cm column of Affi-gel blue gel (Bio-Rad, Woodinville, WA, USA) in 10 mM Tris-HCl buffer (pH 7.4). The unadsorbed proteins (fraction B1) were dialyzed against 10 mM NH4Ac buffer (pH 5) and applied on a 2.5 × 20 cm column of SP-sepharose (GE Healthcare, Uppsala, Sweden). After elution of unadsorbed proteins (fraction S1), the column was eluted with a 0–1 M NaCl concentration gradient in the NH4Ac buffer. The first adsorbed fraction (S2) was then subjected to gel filtration on a Superdex 75 HR 10/30 column (GE Healthcare) in 0.2 M NH4HCO3 buffer (pH 8.5). The second absorbance peak (SU2) represented purified trypsin inhibitor (GBTI).
9
Fenugreek Seed Extraction and Analysis
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Fenugreek (Trigonella foenum-graecum) seeds were purchased from local market.
DEAE-cellulose, glycogen, amylopectin, maltose, pullulan, α-cyclodextrin, β-cyclodextrin, trypsin profile 1GD kit were purchased from Sigma Chemical Co., St. Loius; Mo, U.S.A.
Molecular markers for FPLC were from Pharmacia, Sweden.
All solutions were prepared in Milli Q (Millipore, Bedford, MA, U.S.A.) water.
All the chemicals for buffer were of analytical or electrophoresis grade from Merck Eurolab GmbH Darmstadt, Germany.
DEAE-cellulose, glycogen, amylopectin, maltose, pullulan, α-cyclodextrin, β-cyclodextrin, trypsin profile 1GD kit were purchased from Sigma Chemical Co., St. Loius; Mo, U.S.A.
Molecular markers for FPLC were from Pharmacia, Sweden.
All solutions were prepared in Milli Q (Millipore, Bedford, MA, U.S.A.) water.
All the chemicals for buffer were of analytical or electrophoresis grade from Merck Eurolab GmbH Darmstadt, Germany.
10
Purification and Characterization of LiP
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The crude enzymatic extract was centrifuged at 4000×g for 15 min. The supernatant was firstly brought to 40% saturation by the gradual addition of solid crystals of ammonium sulfate and kept overnight at 4 °C. The precipitate was collected by centrifugation (4000×g) for 15 min at 4 °C and to the supernatant more crystals of ammonium sulfate were added to achieve 80% saturation. It was again kept overnight at 4 °C and centrifuged as described previously. After centrifugation, the sediments were dissolved in sodium tartrate buffer (100 mM l−1, pH 3.5) and dialyzed against the same buffer. The dialysate was applied to DEAE cellulose (Sigma–Aldrich, Brazil) ion-exchange column (3 × 14 cm) which had been equilibrated with sodium tartrate buffer (100 mmol l−1, pH 5.5). The column was washed stepwise with 50, 100, 150, 200, 300 and 500 mmol l−1 de sodium chloride solution (1 mol l−1). The fractions were collected with a flow rate of 1 mL min−1 and the LiP activity and protein contents were determined for each fraction. The active fractions were then pooled and utilized as the partial purified enzymatic extract (PPE) after lyophilization.
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