Metformin, PMSF, leupeptin and aprotinin were purchased from Sigma (St. Louis, MO, USA). Acrylamide, methylene bisAcrylamide, sodium dodecyl sulfonate (SDS), Tris base, ammonium persulfate and Tween-20 were obtained from Amresco (Solon, OH, USA). Fetal bovine serum and DMEM were purchased from HyClone (Logan, UT, USA). Antibodies against p62, phospho-AMPKα (Thr172), phospho-mTOR (Ser2448), mTOR, phospho-p70 S6 Kinase (Thr421/Ser424), p70 S6 Kinase and β-actin were obtained from Cell Signaling (Danvers, MA, USA). Antibodies against PARP1, Caspase-9 and Caspase-3 were purchased from Abcam (Cambridge, MA, USA). Antibody against microtubule-associated protein 1 light chain 3 II (LC3-II) was from GeneTex (Irvine, CA, USA). Anti-rabbit/mouse goat immunoglobulin G-horseradish peroxidase (HRP) secondary antibodies were obtained from Santa Cruz (Santa Cruz, CA, USA). PVDF membrane was purchased from Millipore (Billerica, MA, USA). ECL Plus immunoblotting detection kit was from Bio-Rad (Hercules, CA, USA). Other routine reagents were obtained from Zhongshan (Beijing, China).
Acrylamide
Manufactured by Avantor
Sourced in United States
Acrylamide is a chemical compound commonly used in laboratory applications. It is a colorless, odorless, and water-soluble solid. Acrylamide is commonly used as a monomer in the production of polyacrylamide, a polymer with various industrial and scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
Sodium dodecyl sulfate (sds), by Avantor (7 mentions)
Glycine, by Avantor (5 mentions)
Bis acrylamide, by Avantor (4 mentions)
Ammonium persulfate, by Merck Group (4 mentions)
Ammonium persulfate, by Avantor (3 mentions)
N n methylenebisacrylamide, by Avantor (3 mentions)
Tris base, by Avantor (3 mentions)
Tween 20, by Avantor (3 mentions)
Methanol, by Avantor (3 mentions)
Methylene bisacrylamide, by Avantor (2 mentions)
Hmgb1, by Merck Group (2 mentions)
Hmgb1 elisa kit, by R&D Systems (2 mentions)
Temed, by Avantor (2 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by BD (2 mentions)
Glycerol, by Avantor (2 mentions)
Iodoacetamide, by Bio-Rad (2 mentions)
Bio lyte, by Bio-Rad (2 mentions)
Ethyl pyruvate, by Merck Group (2 mentions)
Bromophenol blue, by Avantor (2 mentions)
Ethylene diamine tetraacetic acid (edta), by Avantor (2 mentions)
15 protocols using acrylamide
1
Immunoblotting Analysis of Autophagy and Apoptosis
2
HMGB1 Isolation and Dilution Protocol
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HMGB1 was purchased from Sigma (USA). The stock solution was aliquoted prior to storage to minimize freezing-thawing cycles. According to the instructions, we obtained the final concentration of 3.7 mg/mL HMGB1 mother liquor by adding 270.27 μL double distilled water into the original bottle with 1mg powder. Subsequently, we divided the mother liquor of HMGB1 into 10μL eppendorf tube and stored these in the refrigerator at −80°C. Before the experiment, a 10μL eppendorf tube of HMGB1 was diluted into 1μg/mL HMGB1 with PBS. During the experiment, 100ng/mL HMGB1 solution was finally obtained by dilution. Fetal bovine serum (FBS), DMEM/F12 was obtained from Gibco (USA). HMGB1 ELISA Kit was purchased from R&D systems (USA). Ethyl pyruvate (EP) was purchased from Sigma (USA). CFSE was purchased from BD (USA). Tris, glycine, TEMED, SDS, and acrylamide were obtained from Amresco (USA).
3
Preparation and Characterization of Formulations
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15d-PGJ2 was from Cayman Chemical (Ann Arbor, Michigan, EUA). tert-Butyl hydroperoxide was from Acros Organics (Geel, Antwerp, Belgium). Brilliant Blue R, reduced glutathione (GSH), hexadecyltrimethylammonium bromide (HTAB), N-ethylmaleimide, o-dianisidine dihydrochloride, phenylmethanesulfonyl fluoride, 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB), nitroblue tetrazolium (NBT), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), Trolox, TPTZ (2,4,6-tris(2-pyridyl)-s-triazine), and bisacrylamide were obtained from Sigma-Aldrich (St. Louis, MO, USA). Xylene cyanol, acrylamide, sodium dodecyl sulfate (SDS), glycerol, and hydroxymethyl aminomethane (Tris) were obtained from Amresco (Solon, OH, USA). ELISA kits for the determination of cytokines were obtained from eBioscience (San Diego, CA, USA). Reverse transcriptase (SuperScript® III), Oligo(dT)12-18 primers, Platinum SYBR Green®, and primers were from Invitrogen (Carlsbad, CA, USA). Isoflurane was from Abbott (Chicago, IL, USA). The excipients used for the preparation of the formulation (Polawax®, caprylic/capric triglyceride, and Phenonip®) were obtained from Galena (Campinas, SP, Brazil). All other reagents used were of pharmaceutical grade.
4
Immunoblot Analysis of Mitochondrial Proteins
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SDS, ammonium persulfate, sucrose, acrylamide, methylene bis-acrylamide, mannitol, glycerol and glycine were purchased from Amresco, LLC. Diazoxide, Nycodenz®, urea, thiourea, EDTA and tetramethylethylenediamine were obtained from Sigma-Aldrich (Merck KGaA). Protein quantification kits, immobilized pH gradient (IPG) strips, dithiothreitol (DTT), BIO-Lyte, CHAPS, agarose, bromophenol blue, β-mercaptoethanol, iodoacetamide and PVDF were acquired from Bio-Rad Laboratories, Inc. Anti-cytochrome c oxidase subunit IV (COX IV; cat. no. ab14744), anti-2-oxoglutarate dehydrogenase (OGDH; cat. no. ab137773), anti-NADH dehydrogenase (ubiquinone) flavoprotein 1 (NDUFV1; cat. no. ab203208) and anti-NADH-ubiquinone oxidoreductase 75 kDa subunit (NDUFS1; cat. no. ab169540) antibodies were purchased from Abcam. All reagents were of analytical grade.
5
Tris-based Protein Extraction Protocol
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Tris, octylphenoxypolyethoxyethanol (NP-40), urea, sulfourea, 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), sodium dodecyl sulphate (SDS), acrylamide, N,N′-methylenebisacrylamide, ammonium persulfate, N,N,N′,N′-tetramethylethylene diamine, CBB G-250 and trichloroacetic acid were obtained from Amresco (Solon, OH, USA); phenylmethylsulfonyl fluoride was obtained from Sigma (St. Louis, MO, USA); immobilized pH gradient strips (IPG strips, 17 cm, pH 4–7) and iodoacetamide were obtained from Bio-Rad Laboratories (Hercules, CA, USA); acetone, glycerol, phosphoric acid, carbinol and alcohol (analytical reagents) were obtained from manufacturers in China. All water used in this experiment was Milli-Q hyperpure water (Millipore, Billerica, MA, USA).
6
Polyacrylamide Gel Electrophoresis Optimization
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Acrylamide (40% w/v) and Bis(Acrylamide) (2% w/v) solutions were bought from Amresco and used without further purification. Tetramethylethylenediamine (TEMED), ammonium persulfate (APS), and sodium acrylate were purchased from Sigma-Aldrich and used as received. Sodium polystyrenesulfonate (NaPSS) of five molecular weights (126, 234, 587, 1188, 2270 kDa) was purchased from ScientiÞc Polymers (catalog numbers 624, 625, 626, 628, and 923). The solutions were prepared using deionized water and sodium chloride. The Hydrophilic Polyvinylidene Fluoride (PVDF) filters with two pore sizes 220 and 450 nm were purchased from Millex Company.
7
Synthesis of Tunable Polyacrylamide Hydrogels
8
Mass Spectrometry Proteomics Sample Preparation
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Tris-base, ammonium persulfate, sodium dodecyl sulfate (SDS), N,N,N’,N’-tetramethylethylenediamine (EDTA), sodium bicarbonate (NH4HCO3), glycine, agarose, urea trichloroacetic acid (TCA), and formic acid were purchased from Sigma (St. Louis, USA). Bio-lyte was purchased from Bio-Rad (Hercules, CA, USA). Acrylamide, N, N’-methylenebisAcrylamide, bromophenol blue, coomassie brilliant blue G-250, thiourea, 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), glycerol, and bovine serum albumin were purchased from Amresco (Solon, OH). Dithiothreitol (DTT) and iodoacetamide were purchased from Merck (Darmstadt, Germany). Trypsin was purchased from Roche (Basel, Switzerland), and trifluoroacetic acid and acetonitrile were from J. T. Baker (Phillipsburg, NJ). Lastly, dipotassium phosphate (K2HPO4), monopotassium phosphate (KH2PO4), sodium chloride (NaCl), sodium hydroxide (NaOH), and acetone were purchased from Beijing Shiji Co. (Beijing, China).
9
SDS-PAGE Reagent and Equipment
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Dulbecco's modified Eagle medium: Nutrient Mixture F-12 (DMEM/F12) (1:1) was purchased from HyClone (Logan, Utah, USA). Fetal bovine serum was obtained from Sijiqing (Hangzhou, China). Gelatin, acrylamide, N,N'-methylenebisacrylamide, glycine, sodium dodecyl sulfate, Triton TM X-100, ethylenediaminetetraacetic acid, Coomassie Brilliant Blue R-250, and sodium azide were from AMRESCO (Solon, OH, USA). Pre-stained protein markers were purchased from Fermentas Co. (Beijing, China). Other reagents were all manufactured domestically to analytical purity. Vertical electrophoresis tanks and the gel imaging apparatus were from Bio-Rad (Hercules, CA, USA).
10
HMGB1 Inhibitor Evaluation Protocol
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HMGB1 was purchased from Sigma(USA).HMGB1 was dissolved in double distilled water(DDW) to make a stock solution(3.7mg/ml) and stored at -80℃.Fetal bovine serum (FBS), DMEM/F12 were obtained from Gibco (USA). HMGB1 ELISA Kit was purchased from R&D systems (USA). Ethyl pyruvate(EP) which is HMGB1 inhibitor was purchased by Sigma(USA).CFSE was purchased from BD(USA).Tris, glycine, TEMED, SDS, acrylamide were obtained from Amresco (USA).
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