BCA PROTEIN ASSAY KIT,MEMBRANE AND CYTOSOL PROTEIN EXTRACTION KIT,TRYPSIN–EDTA SOLUTION,BEYOCOLOR PRESTAINED COLOR PROTEIN MARKER,SDS-PAGE SAMPLE LOADING BUFFER,BEYOECL MOON were purchased from BEYOTIME.PHOSPHO-AMPKa (Thr172) RABBIT mAb and AMPKa ANTIBODY were acquired from CST.STREPTOZOCIN,PVDF MEMBRANCRS,PROTIEN G–AGAROSE, FOETAL BOVINE SERUM and COLLAGENASE were obtained from SIGMA.PERCOOL, SODIUM CITRATE BUFFER and PROTEINASE K were purchased from SOLARBO.GLUT3 ANTIBODY was procured from SCB.2-NBDG was bought from THERMOFISHER.AICAR was acquired from SELLECK.CN.HRP-CONJUGATED AFFNIPURE GOAT ANTI-MOUSE IgG(H + L),HRP-CONJUGATED AFFNIPURE GOAT ANTI-RABBIT IgG(H + L),FITC-CONJ UGATED AFFNIPURE GOAT ANTI-RABBIT IgG(H + L),R-PE-CONJUGATED GOAT ANTI-RABBIT IgG(H + L),CYTOKERATIN 7-SPECIFIC POLYCLONAL ANTIBODY, AMPK ALPHA MONOCLONAL ANTIBODY and ALPHA TUBULIN POLYCLONAL ANTIBODY were purchased from PROTEINTECH.ACRYL/BIS 30% SOLUTION, 1MTRIS- HClSOLUTION(pH6.8),4XTRIS-HCL/SDS(pH8.8),TEMED,TRIS–GLYCINE-SDSRUNNING BUFFER,EZ-BUFFERS C 10X WESTERN TANSFER BUFFER and EZ-BUFFERS H 10X TBST BUFFER were acquired from SANGON BIOTECH. PRIMSCRIP RT MASTER MIX and TB GREEN PREMIX EX TAQ were obtained from TAKARA.
Temed
Manufactured by Sangon
Sourced in China
TEMED (Tetramethylethylenediamine) is a colorless liquid used as a polymerization accelerator in gel electrophoresis applications. It is a commonly used component in the preparation of polyacrylamide gels.
Automatically generated - may contain errors
Lab products found in correlation
Glycine, by Sangon (2 mentions)
Acrylamide, by Sangon (2 mentions)
Collagenase, by Merck Group (1 mentions)
Alpha tubulin polyclonal antibody, by Proteintech (1 mentions)
Sds page sample loading buffer, by Beyotime (1 mentions)
2 nbdg, by Thermo Fisher Scientific (1 mentions)
Aicar, by Selleck Chemicals (1 mentions)
Ripa lysis buffer, by Sangon (1 mentions)
Antibody against nrf2, by Abcam (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Sangon (1 mentions)
Loading buffer, by Sangon (1 mentions)
Gsh px, by Nanjing Jiancheng (1 mentions)
Bca kit, by Sangon (1 mentions)
Bovine serum albumin (bsa), by Sangon (1 mentions)
Nonfat dried milk, by Sangon (1 mentions)
Ammonium sulfate, by Sangon (1 mentions)
Catalase, by Sangon (1 mentions)
Kanamycin sulfate, by Sangon (1 mentions)
5 aminolevulinic acid, by Merck Group (1 mentions)
Ampicillin sodium, by Sangon (1 mentions)
5 protocols using temed
1
Comprehensive Protein Analysis Techniques
2
Seleno-Aminopolysaccharide Antioxidant Pathway
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DAO, D-LA, and LPS kits were obtained from Nanjing Senbeijia Biological Technology Co., Ltd. (Nanjing, China). NO, TNOS, SOD, GSH-Px, CAT, and MDA were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). 2000 DNA Marker, RNA loading buffer, DNA 6×loading buffer, RIPA Lysis buffer, PMSF, BCA kit, BeyoECL Star kit, 5×loading buffer, BSA, non-fat dried milk, TEMED were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Monoclonal antibodies against β-actin were from Cell Signaling Technology, Inc. (Boston, MA, USA). Antibody against Nrf2 was acquired from Abcam (Cambridge, UK). The antibody against Keap1 was acquired from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Alcian Blue, periodic acid schiff were purchased from Beijing solab technology co., Ltd. (Beijing, China). All other reagents are of the highest grade or analytical grade. Our laboratory synthesized the low molecular seleno-aminopolysaccharide (LSA).
3
Cholesterol Oxidation Enzymatic Assay
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All solvents and reagents were of the highest purity commercially available, unless noted otherwise. Isopropyl-β-d -thiogalactopyranoside (IPTG), acrylamide, N,N′-methylenebisacrylamide, ammonium persulfate, 30% acrylamide, TEMED, glycine, Tris, DTT, ampicillin sodium, kanamycin sulfate, chloramphenicol, hydroxypropyl NADH, NADPH, glucose-6-phosphate, glucose-6-phosphate dehydrogenase, catalase and ammonium sulfate were all purchased from Sangon Biotech (Shanghai, China). Cholesterol, Cholesterol propionate, NADPH, 5-aminolevulinic acid, spinach ferredoxin (spFDX) and spinach ferredoxin reductase (spFDR) were obtained from Sigma-Aldrich (St. Louis, MO, USA). 4-Cholesten-3-one and methyl-β-cyclodextrin were purchased from J&K Scientific Ltd. (Beijing, China). Taq DNA polymerase, PrimeSTAR HS DNA Polymerase (with GC buffer), T4 DNA ligase, restriction endonucleases Nde I and Xho I, T-A cloning kits and DNA MW markers were purchased from Takara (Dalian, China). Cholesterol sodium sulfate was purchased from Ark Pharm Inc. (Libertyville, IL, USA).
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
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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