Bowman-Birk inhibitor (BBI) isolated from soybean (Glycine max) was purchased from Sigma-Aldrich (St. Louis, MO). BBI consists of 90% protein as assayed by Biuret, with the remainder of a phosphate buffer salt. The stock solution of BBI was prepared in sterile culture grade water at the concentration of 1 mg/ml. Polybrene was purchased from Sigma-Aldrich (St. Louis, MO). PE-cy7 anti-human CD4 (clone: SK3), FITC anti-human CD195 (CCR5), PE anti-human CXCR4 antibodies for flow cytometry were purchased from BD Bioscience (San Jose, CA), eBiosciences (San Diego, CA), BD Bioscience (San Jose, CA), respectively. The ELISA kits for RANTES, MIP-1α and MIP-1β were purchased from Biolegend Inc. (San Diego, CA), Raybiotech Inc. (Norcross, GA), and Raybiotech Inc. (Norcross, GA), respectively. The neutralization antibodies to RANTES, MIP-1α, MIP-1β were purchased from R & D Systems Inc. (Minneapolis, MN).
Glycine max
Manufactured by Merck Group
Sourced in Germany, United States
Glycine max is a type of lab equipment used for the cultivation and analysis of the soybean plant. It serves as a tool for researchers and scientists to study the properties, growth, and characteristics of the Glycine max species.
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Lab products found in correlation
Mip 1α, by R&D Systems (1 mentions)
Mip 1β, by R&D Systems (1 mentions)
Mip 1β, by BioLegend (1 mentions)
Rantes, by R&D Systems (1 mentions)
Polybrene, by Merck Group (1 mentions)
Neuraminidase, by Merck Group (1 mentions)
Chymotrypsin, by Merck Group (1 mentions)
Vibrio cholerae, by Merck Group (1 mentions)
Trypsin, by Merck Group (1 mentions)
Pisum sativum, by Merck Group (1 mentions)
Triticum vulgaris, by Merck Group (1 mentions)
Phaseolus vulgaris, by Merck Group (1 mentions)
Lycopersicon esculentum, by Merck Group (1 mentions)
Acetylcholinesterase, by Merck Group (1 mentions)
Sodium dodecyl sulphate (sds), by Merck Group (1 mentions)
Linoleic acid, by Merck Group (1 mentions)
Acetylthiocholine iodide, by Merck Group (1 mentions)
Ferrous sulfate, by Merck Group (1 mentions)
Sodium nitrite, by Merck Group (1 mentions)
Eserine, by Merck Group (1 mentions)
6 protocols using glycine max
1
Chemokine Production and Inhibition Assay
2
Enzyme Treatment of Enriched Reticulocytes
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Enriched reticulocytes were prepared with up to 50% hematocrit. Then, the erythrocytes were washed with 500 μl of incomplete RPMI 1640 medium twice by centrifugation at 500 ×g for 3 min at 4°C. Then, the erythrocytes were treated with either neuraminidase (100 mU; from Vibrio cholerae, Sigma-Aldrich Co.), trypsin (0.5 mg; from bovine pancreas, Sigma-Aldrich Co.) or chymotrypsin (0.5 mg; from bovine pancreas, Sigma-Aldrich Co.) at 37°C on a rotator for 1 hr. After enzyme treatment, chymotrypsin- and trypsin-treated RBCs were incubated with a trypsin inhibitor from soybean (Glycine max) (Sigma-Aldrich Co.) at 37°C for 10 min and subsequently washed three times with 10 ml of incomplete RPMI 1640 medium. Packed cells were prepared at a concentration of 1 × 106 cell/ml and used for flow cytometry analysis.
3
Lectin Sourcing and Preparation
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Lectins from: Pisum sativum (L5380), Arachis hypogaea (L0881), Triticum vulgaris (L9640), Glycine max (L1395), Phaseolus vulgaris (61764), Agaricus bisprous (L5640), Lycopersicon esculentum (L2886) were purchased from Sigma-Aldrich, dissolved in sterile phosphate-buffered saline (PBS), and stored at 4°C in a concentration of 1 mg/mL. Succinyl-WGA (W0110) and wheat germ agglutinin FITC-conjugate (L4895), were purchased at Vector Laboratories and Sigma-Aldrich, respectively. These variants were also dissolved in sterile phosphate-buffered saline (PBS) and stored at 4°C in a concentration of 1 mg/mL. Lectin from Sambucus nigra (ZB0106) was purchased from Vector Laboratories. Detailed information on each lectin is included in Table 1 and obtained from Sigma-Aldrich product sheets.
4
Protease Inhibitor Activity Assay
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Extraction of proteins from 100‐ to 150‐mg samples and analysis of PI activity followed a previously described method (Tremacoldi and Pascholati 2002 ; Sarmento et al. 2011 ) with BApNA (Nα‐benzoyl‐DL‐arginine‐4‐nitroanilide hydrochloride, Sigma‐Aldrich, Israel) as the substrate (Mueller and Weder 1989 ). Absorption of the extracts was measured at 405 nm; extraction buffer with or without standard soybean trypsin (Glycine max, Sigma‐Aldrich; 0.1 mg/mL in 1 mmol/L HCl) was used as a control. Based on Thaler et al. (1996 ), PI activity was calculated as the percentage of trypsin activity relative to controls (Thaler et al. 1996 ).
5
Antioxidant and Acetylcholinesterase Assays
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Linoleic acid was purchased from Merck (Darmstadt),. xylenol orange from Searle (England), sodium carbonate from Holpro Analytic (South Africa). Foetal calf serum (FCS), penicillin/streptomycin/fungizone (PSF) and Dulbecco's modified Eagle's medium (DMEM) was obtained from Highveld Biological ( South Africa). Phosphate buffered saline (PBS) and trypsin were purchased from Whitehead Scientific (South Africa). Quercetin, 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide (MTT). sodium dodecyl sulphate, bovine serum albumin (BSA), sodium chloride (NaCl), MgCl 2 •6H 2 O, acetylthiocholine iodide (ATCI), eserine, 5,5dithiobis-2-nitrobenzoic acid (DTNB), acetylcholinesterase (AChE) enzyme from electric eels (type VI-S lypophilized powder), sodium nitrite, ferrous sulfate, indomethacin and 15lipoxygenase from Glycine max purchased from Sigma (Germany) and.
Tris(hydroxymethyl)aminomethane from Sigma, (Switzerland).
Tris(hydroxymethyl)aminomethane from Sigma, (Switzerland).
6
Preparation of Phosphatidylserine Liposomes
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PtdSer derived from bovine brain or soybean (Glycine max; purity ≥ 98％) and phosphatidylcholine (PtdCho) (purity ≥ 98％; both from Sigma-Aldrich, St. Louis, MO, USA) were dissolved in chloroform/methanol (90:10, v/v) . PSL was prepared from a lipid mixture of PtdSer (14 mM) and PtdCho (33 mM) at a molar ratio of 3:7, with or without the fluorescent dye, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(carboxyfluorescein) ( ammonium salt; Avanti Polar Lipids, Inc., Alabaster, Alabama, USA) . The solvent was removed under nitrogen with stirring and dried in a desiccator overnight. The liposomal pellet was then resuspended in PBS (10 mg/mL) .
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