Polyacrylamide (PA) hydrogels were manufactured as described before [17] (link) [18] (link). Briefly, the solutions of 1 kPa PA gel was prepared by mixing 6% acrylamide (Sangon Biotech, China) and 0.05% bis-acrylamide (Sangon Biotech, China), and the solutions of 20 kPa PA gel was prepared by mixing 8% acrylamide and 0.264% bis-acrylamide. Polymerization was initiated with 0.1% tetramethylethylenediamine (TEMED, Klamar, China) and 0.01% ammonium persulfate solution. To make a substrate suitable for each well in a 6-well culture dish, 200 µl of the final solution was dropped onto a glass slide pre-treated with gel slick (Lonza, Switzerland), and a silanized coverslip 30 mm in diameter was placed on the top of the solution. The PA gel yielded was 30 mm in diameter and 200 μm in thickness. After 10 min of polymerization, the glass slide was removed and the coverslip with the gel attached was treated with sulfo-SANPAH (ProteoChem, USA) under UV for 15 min and coated with 0.1 mg/ml rat collagen I (Corning, USA). Before cell seeding, the gels were sterilized by UV for 30 minutes and 75% ethanol.
Acrylamide
Manufactured by Sangon
Sourced in China
Acrylamide is a colorless, odorless, and water-soluble solid chemical compound used in various laboratory applications. It is commonly used as a monomer for the production of polyacrylamide gels, which are widely utilized in gel electrophoresis techniques for the separation and analysis of biomolecules such as proteins and nucleic acids.
Automatically generated - may contain errors
Lab products found in correlation
Bis acrylamide, by Sangon (3 mentions)
Temed, by Sangon (2 mentions)
Methylene bisacrylamide, by Merck Group (1 mentions)
Sodium dodecyl sulfonate sds, by Merck Group (1 mentions)
Tris hydroxymethyl aminomethane tris, by Merck Group (1 mentions)
Coomassie brilliant blue r 250, by Merck Group (1 mentions)
Ammonium persulfate, by Merck Group (1 mentions)
Bile salt, by Merck Group (1 mentions)
2 4 6 trimethylpyridine, by Merck Group (1 mentions)
Boron trifluoride methanol, by Merck Group (1 mentions)
Bicinchoninic acid bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Pepsin, by Merck Group (1 mentions)
α amylase, by Merck Group (1 mentions)
Pancreatin, by Merck Group (1 mentions)
Gelslick, by Lonza (1 mentions)
Rat collagen 1, by Corning (1 mentions)
Sodium dodecyl sulfate (sds), by Sangon (1 mentions)
Glycine, by Sangon (1 mentions)
Tris base, by Sangon (1 mentions)
Superdex peptide 10 300 gl column, by GE Healthcare (1 mentions)
6 protocols using acrylamide
1
Polyacrylamide Hydrogel Fabrication and Functionalization
2
SDS-PAGE Protein Analysis Protocol
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Spectroscopy grade KBr (Tianjin Jinbeier Tech. Co., Ltd., Tianjin, China), SDS-PAGE standard protein (Shanghai Institute of Biochemistry, Chinese Academy of Sciences, Shanghai, China), acrylamide (Sangong Biotech, Shanghai, China), methylene bisacrylamide (Sigma, St. Louis, MO, USA), trishydroxymethylaminomethane (Tris, Sigma), sodium dodecyl sulfonate (SDS, Sigma), ammonium persulfate (Sigma), Coomassie Brilliant Blue R-250 (Sigma), acetic acid (No. 3 Chemical Reagent Factory, Tianjin, China), methanol (Tian Da Reagent Factory, Tianjin, China), tetramethylethylenediamine (TEMED, Sinopharm Chemical Reagent Co., Ltd., Shanghai, China), horseradish peroxidase (HRP, Guo Yuan Biotechnology Co., Ltd., Shanghai, China), glucono-δ-lactone (Nanjing Chemical Reagent Co., Ltd., Nanjing, China). Unless stated otherwise all reagents were analytical purity quality.
3
Bass Nutritional Composition Analysis
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Live large-mouth bass (25±1 cm in length, 378±27 g in weight) was purchased from Vanguard Market (Wuxi, Jiangsu, China) in May and June, 2018. The bicinchoninic acid (BCA) protein assay kit was purchased from Thermo Fisher Scientifi c (Shanghai, China). Standards for 37 fatty acid methyl ester mixture (C4-C24), 14% boron trifl uoridemethanol, α-amylase (from Bacillus licheniformis, 720 U/mg protein, CAS: 9000-85-5), pepsin (from porcine gastric mucosa, 3706 U/mg protein, CAS: 9001-75-6), pancreatin (from porcine pancreas, CAS: 8049-47-6), bile salts, and 2,4,6-trimethylpyridine were obtained from Sigma--Aldrich (Shanghai) Trading Co., Ltd. (Shanghai, China). The 5,5'-dinitrobis[2-nitrobenzoic acid] (DTNB), ethylenediaminetetraacetic acid (EDTA), urea, acrylamide, and bisacrylamide were purchased from Sangon Biotech (Shanghai) Co., Ltd. (Shanghai, China). All other chemicals used were of analytical grade and were from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).
4
Cell Culture Protocols for Gastric Cancer
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The gastric cell lines BGC823, SGC7901, MKN28, AGS and MGC803 were maintained in RPMI-1640 medium, and the Kato3 cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% FBS. All cells were maintained in an incubator (Shellab, Cornelius, Oregon, USA) at 5% CO2 and 37°C. All cell lines were purchased from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). TRAIL) was purchased from Sigma-Aldrich (St Louis, MO, USA). RPMI-1640, DMEM and fetal bovine serum (FBS) were purchased from HyClone (Logan, Utah, USA). Acrylamide, methylene Acrylamide, tris-base, ammonium peroxydisulfate, TEMED, glycine and SDS were purchased from Sangon Biotech, Inc. (Shanghai, China), and the PVDF membrane and chemiluminescence reagents were purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA).
5
Glycosaminoglycan Characterization Protocol
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Chondroitinase (CSase) ABC from Proteus vulgaris, hyaluronidase from sheep testes, heparinase (Hepase) I, Hepase II, Hepase III, and CSase B from Flavobacterium heparinum, protease from Streptomyces griseus, CS-A from bovine trachea, CS-C from shark cartilage, sodium cyanoborohydride, D2O, Alcian Blue, NaH2PO4 and 2-Aminobenzamide (2-AB) were purchased from Sigma Aldrich (Shanghai, China). Keratan sulfate hydrolase (KSase) was prepared as in previous research [29 (link)]. Hep (200 IU/mg) from porcine intestinal mucosa were provided by Dongying Tiandong Pharmaceutical Co., Ltd (Dongying, China). Unsaturated disaccharides CS standards were purchased from Iduron (Manchester, UK). CS-D from shark fins was purchased from Seikagaku Corp. (Tokyo, Japan). Ethanol, acetone, NaCl, NaOH, trichloroacetic acid, ether, NH4HCO3, sodium acetate and acetic acid were purchased from China National Pharmaceutical Group Co., Ltd. (Shanghai, China). Acrylamide and bis-Acrylamide were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). SuperdexTM Peptide 10/300 GL column was obtained from GE Healthcare Life Sciences (Beijing, China). YMC-Pack PA-G column (250 × 4.6 mm ID) was purchased from YMC (Kyoto, Japan). All other reagents were of the highest quality available.
6
Polyacrylamide Gel Compression Protocol
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The polyacrylamide gel used for cell compression was prepared as described in Matthews et al. and Le Berre et al. with modifications.11 (link),59 (link) Briefly, 18 mm glass coverslips (Sangon, F518211) were treated with 10 μL Binding-silane (Sangon, C500226) for 10 min and then were rinsed with 100% ethanol and air-dried. To prepare soft elastic gels (2 kPa), 1 mL polyacrylamide gel solution was prepared by mixing 125 μL 40% w/v acrylamide (Sangon), 35 μL 2% bis-arcylamide (Sangon), 10 μL APS (10% in water, Sangon), and 830 μL water. After adding and mixing 1 μL TEMED (Sangon) into the gel solution, about 350 μL of the final gel solution was immediately transferred onto a flat glass slide and covered by the coverslip pre-treated with the Binding-silane. After polymerization for 20 min, the gel and the attached coverslip were gently removed from the glass slide using a surgical blade and were soaked in PBS for at least 2 h and followed by incubation in cell culture media for overnight. In experiments that involved drug treatment, the gels were incubated in media containing the drugs for overnight.
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