The DNA pull-down assay was carried out as previously described (Liu et al., 2017 (link)). Synthetic cecropin B peptide (cec B) was covalently crosslinked with TANBead USPIO-101 (Taiwan Advanced Nanotech Inc., Taiwan) through a 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (Sigma-Aldrich, USA) by coupling reaction in the coupling buffer (100 mM 2-(N-morpholino)-ethanesulfonic acid, 150 mM Sodium chloride, pH 6.0) and then stored at 4°C until use. The cec B peptide-conjugated magnetic beads were incubated with each PPO 3 DNA fragment for 30 min at 25°C under gentle rotation. The bead-cec B peptide-DNA fragment complexes were then collected with a magnet and was washed three times with 1× cold PBS containing 0.1% Tween-20, followed by two times with 1x cold PBS. Each bead-cec B peptide-DNA fragment complex fraction was then collected again with a magnet and suspended in 50 μL distilled water for subsequent qPCR analysis. The specific primer pairs used to detect the PPO 3 DNA fragments are shown in Table S1 .
1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Italy, Spain, China
1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride is a water-soluble carbodiimide compound commonly used as a crosslinking and activating agent in various chemical and biological applications.
Automatically generated - may contain errors
Lab products found in correlation
N hydroxysuccinimide, by Merck Group (45 mentions)
Bovine serum albumin (bsa), by Merck Group (7 mentions)
Hydrochloric acid (hcl), by Merck Group (7 mentions)
Sodium hydroxide (naoh), by Merck Group (7 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (7 mentions)
Ethanolamine, by Merck Group (6 mentions)
N hydroxysuccinimide nhs, by Merck Group (6 mentions)
N n methylenebisacrylamide, by Merck Group (6 mentions)
Tween 20, by Merck Group (5 mentions)
N hydroxysulfosuccinimide, by Merck Group (5 mentions)
2 n morpholino ethanesulfonic acid, by Merck Group (5 mentions)
Phosphate buffered saline (pbs), by Merck Group (5 mentions)
Acetonitrile, by Merck Group (5 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Merck Group (4 mentions)
Dicyclohexylcarbodiimide, by Merck Group (4 mentions)
Pyridine, by Merck Group (4 mentions)
Glutaraldehyde, by Merck Group (4 mentions)
Sodium chloride (nacl), by Merck Group (4 mentions)
Ammonium persulfate, by Merck Group (4 mentions)
N tert butylacrylamide, by Merck Group (4 mentions)
99 protocols using 1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride
1
DNA-Binding Peptide Pulldown Assay
2
Synthesis of Nanomaterial-based Immunosensors
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IAA, salicylic acid (SA), abscisic acid (ABA), gibberellic acid (GA), potassium ferricyanide (K3[Fe(CN)6]), chloroauric acid (HAuCl4) and trisodium citrate were purchased from Aladdin (Shanghai, China). 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), 4-aminophenylboronic acid (APBA), rat monoclonal antibody against IAA (anti-IAA) and 11-mercaptoundecanoic acid (MUA) were provided by Sigma (U.S). 2,5-Dimercapto-1,3,4-thiadiazole (DMcT) was the product of Alfa Aesar (U.S). Goat anti-rat immunoglobulin G (IgG) was purchased from Sangon Co., Ltd. (Shanghai, China). MWCNTs of ca. 20–40 nm average diameter were purchased from Shenzhen Nanotech Port Co., Ltd. (Shenzhen, China). Aniline, ammonium persulfate (APS) and ethylene glycol were purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Phosphate buffer solution (PBS) was prepared by mixing the stock solutions of 0.1 M NaH2PO4 and 0.1 M Na2HPO4, and the pH was adjusted using NaOH or HCl. Double-distilled deionized water was used throughout the experiments.
AuNPs,13 (link) IgG–AuNPs,13 (link) and anti-IAA-AuNPs10 (link) were synthesized according to the previous reports.
AuNPs,13 (link) IgG–AuNPs,13 (link) and anti-IAA-AuNPs10 (link) were synthesized according to the previous reports.
3
Synthesis of Fluorescent Silica Nanoparticles
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Example 1
All the chemicals and reagents, including cetyltrimethylammonium bromide (CTAB, ≥98%, Sigma-Aldrich), sodium hydroxide (AG, Fluka), tetraethyl orthosilicate (TEOS, ≥99%, Sigma-Aldrich), 3-aminopropyl triethyl silane (APTS, 99%, Sigma-Aldrich), 1-pyrene carboxylic acid (97%, Sigma-Aldrich), hydroxy benzotriazole (≥97%, Sigma-Aldrich), 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide hydrochloride (≥99%, Sigma-Aldrich), triethylamine (≥99%, Sigma-Aldrich), ethyl acetate (High-performance liquid chromatography (HPLC), Honeywell) were purchased and utilized without further purification.
4
Multifunctional Nanoparticle Delivery System
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Oridonin (> 98%) was purchased from ToYongBio (Shanghai, China). Poly(D,L-lactide-coglycolide)-b-poly(ethylene glycol)-carboxylic acid (5 k - 20 k) was purchased from Xi’an Qiyue Biology (Xi’an, Shanxi Province, China). Coumarin 6 (98%), Evans blue dye (≥ 75%), formamide, lipopolysaccharides (LPS, Escherichia coli O55:B5 in vivo and Escherichia coli O111:B4 in vitro), dichloromethane (DCM), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-hydroxysulfosuccinimide (NHS), hexadecyltrimethylammonium bromide (HTAB, ≥ 98%), O-dianisidine dihydrochloride (O-DHC) and PVA (1%, w/w) were purchased from Sigma-Aldrich (St. Louis, MO, USA). MES hydrate (99+%) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Purified anti-mouse CD31 antibody was purchased from Biolegend (San Diego, CA, USA). Primary antibodies against VE-cadherin, Nrf2, HO-1, Keap-1, p62, and β-actin and siRNA p62 were purchased from Cell Signaling Technology (Boston, MA, USA). Phalloidin was purchased from Cytoskeleton, Inc. (Denver, CO, USA). Fluorescence-conjugated secondary antibodies (Alexa Fluor® 488) were purchased from Abcam (Cambridge, UK).
5
PLGA Nanoparticles for Targeted Drug Delivery
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Poly(D, L -lactic-co-glycolic acid) (PLGA, with a lactide/glycolide molar ratio of 75:25 and an inherent viscosity of 0.17 dL g-1) was obtained from Daigang BIO Engineer Ltd. Co. (Shandong, China). Catalase, H2O2, GSH, 1-ethyl-3-[3-(dimethylamino)propyl] carbodiimidehydrochloride (EDC), N-hydroxysuccinimide (NHS), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl- tetrazolium bromide (MTT), 9,10-phenan-threnequinone, poly(vinyl alcohol) (PVA, MW=57-66 kDa), 3,3'-dioctadecyloxacarbocya-nineperchlorate (DiO) and other chemical reagents were all obtained from Sigma-Aldrich (St. Louis, MO, USA). c(RGDfK) was obtained from Sangon Biotech Ltd. Co (Shanghai, China). Hoechst 33342, LysoTracker Red, MitoTracker Red, ER Tracker Red were obtained from Invitrogen (Carlsbad, CA, USA). RA-V was isolated from Rubia yunnanensis as described earlier 6 (link). Ultrapure water was prepared using a Millipore Simplicity System (Millipore, Bedford, USA).
6
LyP-1 Peptide Conjugation and Characterization
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LyP-1 (CC (Acm) GNKRTRGC (Acm)) was purchased from Peptron, Inc. Yuseong-gu, Daejeon, Korea, having 98% HPLC purity. Branched polyethyleneimine (PEI) (MW = 25 KDa), low-molecular-weight chitosan (MW = 50 kDa, DE = 75–85%), human recombinant endostatin (MW = 22 kDa), N-hydroxysuccinimide (NHS), 1-carbonyldiimidazole (CDI), sodium tripolyphosphate (TPP) (MW = 367.86 g/mol), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), poly(vinyl alcohol) (PVA) (MW = 85,000 g/mol), triethylamine dimethylformamide (DMF) and fluorescein isothiocyanate (FITC) (MW = 389.382 g/mol) were all purchased at Sigma-Aldrich (St. Louis, MO, USA) Modified NH2-PEG-COOH (Mw = 2100 g/mol) was procured from NANOCS (New York, NY, USA). Fetal bovine serum (FBS), RPMI, KYSE-30, HAM’s F12 and pentamycin/streptomycin were procured from Life Biosciences (Oakleigh, VIC, Australia) and DAPI (4′,6-diamidino-2-phenylindole) solution was obtained from Thermo Fisher Scientific (Waltham, MA, USA).
7
Diagnostic Assay for Potato Viruses
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Ordinary and necrotic types of potato virus Y (PVYN, PVYO), potato viruses X, M, S, A, and potato leafroll virus were obtained from the A.G. Lorch All-Russian Potato Research Institute (Moscow, Russia). Tris(hydroxymethyl)aminomethane (Tris), Triton X-100, 3,3′,5,5′-tetramethylbenzidine dihydrochloride (TMB), sodium azide, Tween-20, bovine serum albumin (BSA), chloroauric acid, N-hydroxysuccinimide (NHS), 1-ethyl-3(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), biotinamidohexanoyl-6-aminohexanoic acid N-hydroxysuccinimide ester, ethanolamine, and a conjugate of streptavidin-polyperoxidase were obtained from Sigma-Aldrich (St. Louis, MO, USA). Protein A derived from Staphylococcus aureus was purchased from Imtek (Moscow, Russia). All of the salts, acids, and solvents were of analytical reagent or chemical reagent grade. All of the solutions that were used to obtain the GNPs and their conjugates were prepared using Milli-Q water (Millipore, MA, USA).
The nitrocellulose membranes (CNPC-12μ) adhering to the surface of a laminated card, conjugate release matrix (PT-R5), sample pads (GFB-R4, 0.35), and absorbent pads (AP045) were obtained from Advanced Microdevices (Ambala Cantt, Haryana, India).
The nitrocellulose membranes (CNPC-12μ) adhering to the surface of a laminated card, conjugate release matrix (PT-R5), sample pads (GFB-R4, 0.35), and absorbent pads (AP045) were obtained from Advanced Microdevices (Ambala Cantt, Haryana, India).
8
Synthesis and Characterization of DHA-Based Theranostic Nanoparticles
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DHA, 3-mercaptopropionic acid, N-hydroxysuccinimide (NHS), succinic anhydride, 4-dimethylaminopyridine (DMAP), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), and dicyclohexylcarbodiimide were purchased from Sigma-Aldrich. DOTA-NH2 was purchased from Confluore Biological Technology Co. Ltd. Fer-1 and DOX hydrochloride were purchased from MedChemExpress. CDDP and DFO were purchased from Energy Chemical. Anti–4-HNE (bs-6313R) was purchased from Bioss. Anti-GPX4 (ab125066) was purchased from Abcam. Anti-TfR1 (BA0462) was purchased from Boster. Anti–FTH-1 (no. 3998) was purchased from Cell Signaling Technology. Slc40A1 (Fpn, AB-23233-A) was purchased from Alpha Diagnostics. β-Actin (8H10D10) was purchased from Cell Signaling Technology. FerroOrange was purchased from Maokang Biotechnology. The creatinine assay kit and BUN assay kit were purchased from Ponstar Biotech Co. Ltd.
9
Chitosan-Dextran Sulfate Nanoparticles for Lutein Delivery
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Chitosan shrimp (MW 30 kDa with 95% degree of deacetylation) was obtained from Aquapremier Inc. (Bangkok, Thailand). Dextran sulfate (MW 500,000 Da), 1-ethyl-3-(3- dimethylaminopropyl)-carbodiimide hydrochloride and type II mucin from porcine stomach were purchased from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). Polyethylene glycol 400 was purchased from Namsiang Trading Co., Ltd. (Bangkok, Thailand). Lutein (95% lutein extracted from marigold flower extract) was purchased from KEB Biotechnology Co., Ltd. (Beijing, China). All other chemicals and reagents used were of high-performance liquid chromatography (HPLC) and analytical grade. Deionized (DI) water was used in the preparation of solutions and dispersion of nanoparticles.
10
Functionalized Polystyrene Microspheres for CA 19-9 Detection
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Polystyrene microspheres with diameter of 400 nm (10 wt. %) were obtained from Bangs Laboratories Inc., USA. Cysteamine, (3-Aminopropyl)triethoxysilane (APTES), 4arm-Polyethylene glycol-Carboxyl (Mn 10,000), dimethylformamide (DMF), 1-ethyl-3-[3-(dimethylamino)propyl]-carbodiimide hydrochloride (EDC), N-Hydroxysuccinimide (NHS), donkey serum, Dylight800 conjugated polyclonal donkey anti-rabbit antibody and phosphate buffered saline (PBS, pH 7.4) were purchased from Sigma-Aldrich, UK. Nanopure water (>18.2 MΩ), purified using the Millipore Milli-Q gradient system, was used in all the experiments. Glass microscope slides were obtained from VWR International, USA and rinsed with acetone, 2-propranol and nanopure water before use. Purified Carbohydrate antigen (CA 19–9) and monoclonal mouse anti-CA 19–9 antibody were purchased from Biospacific. Polyclonal rabbit anti-CA 19–9 antibody was purchased from Lifespan Biosciences.
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