Kanamycin sulfate, tetracycline hydrochloride, streptomycin, chloramphenicol, gentamicin sulfate were purchased from Adamas Reagent, Ltd. Streptomycin sulfate was purchased from Tokyo Chemical Industry Co., Ltd. The above antibiotics were made into a solution with a concentration of 0.01 M and prepared with ultrapure water kept at 4 °C. The aptamer (FAM–apt, 5′-FAM-TGG GGG TTG AGG CTA AGC CGA-3′) was synthesized at Sangon Biotech (Shanghai) Co., Ltd. Phosphate buffer saline (PBS) and TE (Tris–EDTA) buffer as Oligonucleotide stock solution were purchased from Sangon Biotech (Shanghai) Co., Ltd. and kept at 4 °C. The HNO3 and HCl needed for glassware cleaning are purchased from Yonghua Chemical Co., Ltd., and HNO3 and HCl are prepared in a ratio of 1 : 3. All chemical reagents used are analytical reagents, and all solutions are prepared in autoclaved double distilled deionized water.
Phosphate buffer saline (pbs)
Manufactured by Sangon
Sourced in China
Phosphate buffer saline (PBS) is a commonly used buffer solution in various laboratory applications. It is an aqueous solution containing sodium phosphate and sodium chloride, maintaining a physiological pH and osmolarity. PBS is primarily used to maintain the integrity and stability of biological samples, cells, and proteins during experiments and procedures.
Automatically generated - may contain errors
Lab products found in correlation
Alamethicin, by BD (2 mentions)
Uridine 5 diphosphoglucuronic acid trisodium salt, by BD (2 mentions)
Magnesium sulfate mgso4, by Tianjin Guangfu (2 mentions)
Streptomycin sulfate, by Tokyo Chemical Industry (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
β nicotinamide adenine dinucleotide phosphate, by Merck Group (1 mentions)
Peptone, by Sangon (1 mentions)
Tryptone, by Sangon (1 mentions)
Yeast extract, by Sangon (1 mentions)
Vitamin k1, by Sangon (1 mentions)
L cysteine, by Sangon (1 mentions)
Paraformaldehyde (pfa), by Sangon (1 mentions)
Cel mir 39 3p standard rna, by RiboBio (1 mentions)
Bca protein concentration assay kit, by Boster Bio (1 mentions)
Paraformaldehyde (pfa), by Sinopharm (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
Crystal violet, by Sangon (1 mentions)
12 protocols using phosphate buffer saline (pbs)
1
Optimized Antibiotic Preparation for Aptamer Assays
2
Eriocitrin Biochemical Characterization
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Eriocitrin (13463-28-0, purity > 98.5%) was purchased from Chengdu Desite Co., Ltd. (Chengdu, China). β-Nicotinamide adenine dinucleotide phosphate (NADPH) was purchased from Sigma Chemical (St. Louis, MO, USA). Alamethicin and uridine 5′-diphosphoglucuronic acid trisodium salt (UDPGA) were purchased from BD Biosciences (Woburn, MA, USA). Phosphate buffer saline (PBS) was purchased from Sangon Biotech (Shanghai) Co., Ltd. Acetonitrile, methanol and formic acid were all HPLC grade and were purchased from Fisher Scientific (Waltham, MA, USA). Dilute hydrochloric acid (HCl) was purchased from Shijiazhuang Reagent Factory. Purified water was purchased from Wahaha (Hangzhou Wahaha Group Co., Ltd.). l -Ascorbic acid, l -cysteine, eurythrol, tryptone and nutrient agar were purchased from Beijing AoBoXing Bio-tech (Beijing) Co., Ltd. Sodium carboxymethyl cellulose (CMC–Na), sodium carbonate (Na2CO3), magnesium chloride (MgCl2), potassium dihydrogen phosphate (KH2PO4), dipotassium phosphate (K2HPO4), calcium chloride (CaCl2), ammonium sulfate ((NH4)2SO4), sodium chloride (NaCl) and magnesium sulfate (MgSO4) were obtained from Tianjin Guangfu Technology Development Co., Ltd. (Tianjin, China).
3
Fluorescence Probes for Cell Imaging
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FDAA probes were bought from Chinese Peptide Company (Hangzhou, China). FISH probes used in this study were synthesized and labeled at the 5′ ends with FAM (carboxyfluorescein) by Sangon Biotech (Shanghai, China). Tryptone, peptone, calf brain infusion, beef heart infusion, yeast extract, glucose, L-cysteine, vitamin K1, vitamin K3, paraformaldehyde (PFA) and phosphate buffer saline (PBS) were purchased from Sangon Biotech (Shanghai, China). Other chemicals, not noted above, were from Sigma–Aldrich (St. Louis, MO, USA).
4
Eupatorin Compound Characterization and Synthesis
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Eupatorin (855-96-9, purity > 98.94%) was purchased from Chengdu Desite Co., Ltd. (Chengdu, China). Beta-nicotinamide adenine dinucleotide phosphate (β-NADPH) was purchased from Sigma Chemical (St. Louis, MO, USA). Alamethicin and uridine 5′-diphosphoglucuronic acid trisodium salt (UDPGA) were purchased from BD Biosciences (Woburn, MA, USA). Phosphate buffer saline (PBS) was purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Acetonitrile and methanol were all HPLC grade and were purchased from J.T.-Baker Company (Phillipsburg, NJ, USA). Formic acid (HPLC grade) was provided by Diamond Technology (Dikma Technologies Inc., Lake Forest, CA, USA). Purified water was purchased from Wahaha (Hangzhou Wahaha Group Co., Ltd., Hangzhou, China). L-ascorbic acid, L-cysteine, eurythrol, tryptone and nutrient agar were purchased from Beijing AoBoXing Bio-tech Co., Ltd. (Beijing, China). Sodium carboxymethyl cellulose (CMC-Na), sodium carbonate (Na2CO3), magnesium chloride (MgCl2), potassium dihydrogen phosphate (KH2PO4), dipotassium phosphate (K2HPO4), calcium chloride (CaCl2), ammonium sulfate ((NH4)2SO4), sodium chloride (NaCl) and magnesium sulfate (MgSO4) were obtained from Tianjin Guangfu Technology Development Co., Ltd. (Tianjin, China).
5
Reagents and Materials for Molecular Experiments
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In this experimental study, phosphate buffer saline (PBS) was purchased from Sangon
Biotech Co., Ltd (Shanghai, China) Paraformaldehyde (4%) was obtained from China National
Pharmaceutical Group Corporation (Shanghai, China). The BCA Protein Concentration Assay
Kit was purchased from Boster Biological Technology Co. Ltd (Wuhan, China). RNAiso Plus
(TRIzol) and PrimeScriptTM II 1st Strand cDNA Synthesis Kit were purchased from
Takara Biomedical Technology (Beijing) Co., Ltd (Beijing, China). The cel-mir-39-3p
standard RNA was obtained from Guangzhou RiboBio Co., Ltd (Guangzhou, China).
Biotech Co., Ltd (Shanghai, China) Paraformaldehyde (4%) was obtained from China National
Pharmaceutical Group Corporation (Shanghai, China). The BCA Protein Concentration Assay
Kit was purchased from Boster Biological Technology Co. Ltd (Wuhan, China). RNAiso Plus
(TRIzol) and PrimeScriptTM II 1st Strand cDNA Synthesis Kit were purchased from
Takara Biomedical Technology (Beijing) Co., Ltd (Beijing, China). The cel-mir-39-3p
standard RNA was obtained from Guangzhou RiboBio Co., Ltd (Guangzhou, China).
6
Transwell Invasion and Migration Assay
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A total of 40 μL Matrigel (BD biosciences, San Jose, CA, USA) at the dilution of 1:8 was added to the transwell upper chambers for 30 minutes at 37°C for solidification to conduct transwell invasion assay, while un-coated transwell chambers were used for transwell migration assay. After transfection for 24 hours, pancreatic cancer cells in serum-free medium were seeded into the upside of the membrane, while the lower chambers were filled with medium added with 10% FBS. Transwell plates were placed in a cell incubator for 24 hours. Cells migrated or invaded to the lower surface of the membrane were washed with phosphate buffer saline (PBS, Sangon Biotech, Shanghai, People's Republic of China), fixed with 4% paraformaldehyde (Sigma, St. Louis, MO, USA) and stained with 0.5% crystal violet (Sangon Biotech). The numbers of migrated and invaded cells in five random fields were counted at 100 × magnification. The scale bar indicates 200 μM.
7
Plasmonic Nanomaterial Synthesis and Biofunctionalization
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Gold chloride tetrahydrate (HAuCl4), tris(2-carboxyethyl) phosphonate hydrochloride (TCEP), dichlorotris(1, 10-phenanthroline) ruthenium hydrate (Ru(phen)3Cl2·H2O), triethylamine (TEA) with other analytical grade chemicals were obtained by Aladdin Biochemical Technology Co. Ltd. (Shanghai, China). Silver nitrate (AgNO3) and Ti3C2 were purchased from Boer Chemical Reagent Co. Ltd. (Shanghai, China) and XFNANO (Nanjing, China), respectively. L -ascorbic acid (AA) was from Sigma-Aldrich Co. Ltd. (Shanghai, China). T4 DNA ligase and buffer reagents consisting of tris-borate-EDTA (TBE) and phosphate buffer saline (PBS) were obtained from Sangon Biotechnology Co. The purified water in the experiments was acquired by the Milli-Q purification system (Branstead), and the resistance value was kept above 18 MΩ. The oligonucleotides and modified nucleic acid probes employed in the entire protocol were purchased from Genscript Biotechnology Co. Ltd. (Nanjing, China). The related sequences used in the experiment are depicted in Table S1 .
8
Electrochemical Detection of SARS-CoV-2 Nucleoprotein
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Synthetic oligonucleotides were ordered and HPLC-purified by General biol. Co., Ltd. (Chuzhou, China), and the sequences were listed in Table S1 . The aptamers used in this study were developed by our group [20] . Zirconium (IV) chloride (ZrCl4) and amino-terephthalic acid (NH2-BDC) were purchased from Heowns Chemical Reagent (Tianjin, China). Gold chloride tetrahydrate (HAuCl4·4H2O) was obtained from Sigma-Aldrich (St. Louis, MO). Phosphate buffer saline (PBS) and bovine serum albumin (BSA) were purchased from Sangon Biotech (Shanghai, China). National center for protein science Shanghai (NCPSS) offered the plasmid of SARS-CoV-2 N protein (NFPS–P10034 Pet28a-N). A Millipore filtration system produced deionized water used throughout this experiment. All chemical reagents were of analytical grade and used without further purification.
SARS-CoV-2 Np was prepared and purified based on our previous procedures [22] , and the purity was checked by SDS-PAGE (Fig. S1 ). A portable potentiostat (Ivium, Netherlands) was operated to record electrochemical signals while screen-printed gold electrodes (SPGEs, ref. DRP-250BT, ϕ = 4 mm) were purchased from Metrohm DropSens S.L. (Spain). A BIAcore T200 biosensor system was used to conduct surface plasmon resonance (SPR) experiments to assess the binding affinity of aptamers.
SARS-CoV-2 Np was prepared and purified based on our previous procedures [22] , and the purity was checked by SDS-PAGE (
9
Antimicrobial Polymer Synthesis and Evaluation
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Ethanol absolute (AR Grade, ≥99.7%) and chloroform (AR Grade, ≥99.0%) were purchased from Sinopharm Chemical Reagent Shanghai Co., Ltd. (Shanghai, China). Acetic acid (AR Grade, ≥99.5%), phosphate buffer saline (PBS buffer), and dimethyl sulfoxide (DMSO, molecular biology grade, ≥99.7%), 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydro (EDC, ≥95.0%, C600433-0025), glucose (≥99.8%, A100188-0005) were purchased from Sangon Biotechnology Co., Ltd. (Shanghai, China). E. coli (ATCC 43888) and S. aureus (ATCC BAA-1721) were purchased from China Center of Industrial Culture Collection, CICC. Chlorhexidine acetate (CHA, ≥99.0%, C107054), dithiothreitol (DTT, ≥99.0%, D104859), cysteamine dihydrochloride (CYS, ≥97.0%, C153647), protocatechuic acid (PCA, ≥97.0%, S30117), polyethylene glycol 1000 (PEG1000, AR Grade, 12803702) were purchased from Aladdin (Co. Ltd. Shanghai, China). MMT k-10 (M813515), 3-carboxyphenyl boric acid (CPBA, ≥99.0%, C804442), 4-Dimethylaminopyridine (DMAP, ≥97.0%, D807273), N-Hydroxysuccinimide (NHS, ≥97.0%, N811124) Ethylene imine polymer (PEI, ≥99.0%, E808878), Deuterium oxide (D2O, ≥99.9%, D807644), Methanol-d4 (CD3OD, ≥99.9%, M812876), DMSO-d6 (≥ 99.9%, D806935) were purchased from Macklin Biochemical Co., Ltd. (Shanghai, China).
10
Caspase-3 Activation and Mitochondrial Dynamics in Brains
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Mice brains were dehydrated with 30% sucrose solution and embedded in a tissue freezing medium (OCT, SAKURA Tissue-Tek, CA, United States). The frozen samples were sectioned for 5 µm thickness and fixed with 50% ethanol, including 5% glacial acetic acid and 5% formaldehyde. After washing in phosphate buffer saline (Sangon Biotech, Shanghai, China), brain samples were blocked in 5% goat serum at 37°C for 20 minutes. The samples were incubated with primary antibodies [p-DRP1 (1:800, #4494, Cell Signaling Technology, Boston, MA), DRP1 (1:50, #8570, Cell Signaling Technology, Boston, MA), cleaved-caspase 3 (1:400, #9661, Cell Signaling Technology, Boston, MA)], Dihydroethidium (DHE) and NeuN at 4°C overnight. Tissues were incubated with secondary antibodies (1:200, #A1034, Thermo Fisher Scientific, CA; 1:200, #115–585–003, Jackson, MI) at 37°C for 1 hour. Images were obtained using a fluorescence microscope (Olympus, Tokyo, Japan).
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