Poly[dimethylsiloxane-co-(3-aminopropyl)methylsiloxane] with a functional group equivalent weight of 4400 Da (silicone oil), N,N′-diisopropylcarbodiimide (DIC), N,N′-dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole hydrate (HOBt), 1,1′-carbonyldiimidazole (CDI), 3-mercaptopropionic acid (MPA), l -cysteine hydrochloride monohydrate (cysteine), 4,4′-dithiodipyridine (DTDP), 2,4,6-trinitrobenzenesulphonic acid solution 5% (w/v) in demineralized water (TNBS), iodine, pyridine, triethylamine and 1-(2-methoxyphenylazo)-2-naphthol (sudan red G) were purchased from Sigma–Aldrich (Steinheim, Germany). All other chemicals, reagents and solvents were received from commercial sources.
L cysteine hydrochloride monohydrate
Manufactured by Merck Group
Sourced in Germany, United States, Italy
L-cysteine hydrochloride monohydrate is a chemical compound used as a laboratory reagent. It is a crystalline solid that is soluble in water and has a molecular formula of C3H7NO2S·HCl·H2O.
Automatically generated - may contain errors
Lab products found in correlation
Gemini c18 column, by Phenomenex (4 mentions)
Luciferase assay kit, by Promega (2 mentions)
L histidine hydrochloride, by Sangon (2 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (2 mentions)
L arginine, by Sangon (2 mentions)
Thiourea, by Merck Group (2 mentions)
Anaerocult a, by Merck Group (2 mentions)
Yeast extract, by Merck Group (2 mentions)
Dithiothreitol (dtt), by Merck Group (2 mentions)
Sodium borohydride, by Merck Group (2 mentions)
L ascorbic acid, by Merck Group (2 mentions)
Gaspak ez, by BD (2 mentions)
β mercaptoethanol, by Merck Group (2 mentions)
Hydrocortisone, by Merck Group (2 mentions)
N n diisopropylcarbodiimide, by Merck Group (1 mentions)
1 hydroxybenzotriazole hydrate, by Merck Group (1 mentions)
1 1 carbonyldiimidazole, by Merck Group (1 mentions)
3 mercaptopropionic acid, by Merck Group (1 mentions)
4 4 dithiodipyridine, by Merck Group (1 mentions)
2 4 6 trinitrobenzenesulphonic acid, by Merck Group (1 mentions)
35 protocols using l cysteine hydrochloride monohydrate
1
Functionalization of Silicone Oil for Biomedical Applications
2
Optimizing Transfection Efficiency for Gene Delivery
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Materials used in this study were l -histidine hydrochloride, l -arginine, stearic acid, and hydrogen peroxide (30% w/v; Sangon Biotech, Shanghai, People’s Republic of China); polyethylenimine (PEI, branched, molecular weight 25 kDa), Lipofectamine™ 2000, l -buthionine-sulfoximine, 4′,6-diamidino-2-phenylindole dihydrochloride, l -cysteine hydrochloride monohydrate, ethidium bromide, dithiothreitol (DTT, Sigma-Aldrich, St Louis, MO, USA); a luciferase assay kit (Promega, Madison, WI, USA); pDNA (pGL3 and pEGFP) (Shanghai Innovation Biotechnology Co Ltd, Shanghai, People’s Republic of China); enhanced bicinchoninic acid protein assay kit (Beyotime, Nanjing, People’s Republic of China); Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum, YOYO-1 (Y3601) and penicillin–streptomycin solution 5 kU/mL (Life Technologies, Carlsbad, CA, USA); a Cell Counting Kit-8 (CCK-8, Dojindo Molecular Technologies Inc, Nanjing, People’s Republic of China); and HEK293 cells and HeLa cells (Cell Culture Center of the Shanghai Institutes for Biological Sciences of the Chinese Academy of Sciences, Shanghai, People’s Republic of China). All other reagents were of analytical grade. All animal experiments were performed in accordance with the ethics and regulations of animal experiments of Second Military Medical University (Shanghai, People’s Republic of China).
3
Anaerobic Culturing of Oral Bacteria
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The bacterial strains used in this study were Fusobacterium nucleatum ATCC25586 (FN), Actinomyces naeslundii X600 (AN) and Streptococcus mitis ATCC 903 (SM). These bacterial strains were cultured anaerobically in BHIS broth using the AnaeroPack System (Mitsubishi Gas Chemical Company, Inc.). BHIS broth is Brain Heart Infusion medium (Eiken Chemical Co.) supplemented with 5 µg/ml hemin (Sigma-Aldrich), 0.1% L-cysteine hydrochloride monohydrate (Sigma-Aldrich), 0.5% yeast extract (Becton Dickinson and Company) and 0.375% D-glucose (Nacalai Tesque).
4
Mucoadhesive Dexamethasone Delivery
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Hydroxypropyl-β-cyclodextrin (HP-β-CD) MW 1380 Da (SR: 0.6), dexamethasone (DMS), thiourea, fluorescein diacetate (FDA), Ellman’s reagent (2,2′-dinitro-5,5′-dithiobenzoic acid), l -cysteine hydrochloride monohydrate, tris(hydroxymethyl)aminomethane, Sephadex G-15, Type II porcine gastric mucin, Dulbecco’s Modified Eagle Medium (DMEM), calf bovine serum, a mixture of antibiotics consisting of an aqueous solution of penicillin (10,000 U/mL) and streptomycin (10,000 µg/mL), 10 mM phosphate buffer pH 7.3 without Ca2+ and Mg2+ (PBSA), and a trypsin-EDTA buffer solution containing 0.25% trypsin were all purchased from Sigma-Aldrich (Darmstadt, Germany). Fibroblast BALB/3T3 clone A31 cells (CCL-163) were obtained from American Type Culture Collection.
5
PLGA-Based Insulin Delivery System
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PLGA (copolymer composition ratio of 50 : 50, weight average molecular weight of 20 kDa, and inherent viscosity of 0.187 to 0.229 dL/g), methylene chloride (CH2Cl2), polyvinyl alcohol (86–90 mol% hydrolysis), recombinant human insulin, hydrochloric acid (HCl), sodium hydroxide (NaOH), absolute ethanol (99.5%), N-hydroxysuccinimide esters (NHS), 25% glutaraldehyde solution, and sodium dihydrogen phosphate (NaH2PO4) were obtained from Wako Pure Chemicals Ltd., Japan. L-cysteine hydrochloride monohydrate (minimum 98%), ethylene diamine tetra acetic acid (EDTA), papain, DNA quantification kit, Dulbecco's Modified Eagle's Medium (DMEM), growth supplements, and antibiotics were obtained from Sigma-Aldrich, USA. Phosphate buffer saline (10x, pH = 7.4) was obtained from Nacali Tesque Inc., Japan. Porcine collagen type-1 was obtained from Nitta Gelatin, Japan. 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC/EDAC) was obtained from Peptide Institute Inc., Japan. Cellstain Double Staining Kit was obtained from Dojindo Laboratories, Japan. Micro BCA protein assay Kit was obtained from Pierce Biotechnology, USA. All the materials in this study were used as received without further purification. Molecular biology grade milli-Q water from millipore water system (Millipore Corporation, USA) was used for preparation of all the solutions and reagents.
6
Chitosan-based Nanoparticle Drug Delivery
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Chitosan (CS, medium molecular weight, deacetylation degree = 75∼85%), L-cysteine hydrochloride monohydrate (Cys), L-alpha-phosphatidylcholine 14–23% choline basis (PC), and cholesterol were purchased from Sigma‒Aldrich. Analytical grade N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDAC) and N-hydroxysuccinimide (NHS) were purchased from Qi Yun Biotechnology Company (Guangzhou, China). Dox and Cur were obtained from Shuoheng Biotechnology Company (Guangzhou, China). MCF-7 cells were purchased from American Type Culture Collection (ATCC, CRL2593).
7
Integrated Gut Microbiome Modulation
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Formic acid, bile salts, soluble starch, (+)-arabinogalactan, tryptone, yeast extract, xylan from birchwood, L-cysteine hydrochloride monohydrate, guar gum, inulin, Tween 80, buffered peptone water, Dulbecco′s phosphate buffer saline (PBS), casein sodium salt from bovine milk, pectin from citrus fruits, mucin from porcine stomach-type III, CaCl2, KCl, NaCl, NaHCO3, anhydrous K2HPO4, KH2PO4, MgSO4 monohydrate, FeSO4 heptahydrate, resazurin redox indicator, quercetin, kaempferol, quercetin-3-O-rutinoside (aka rutin), phenylacetic acid, 4′-hydroxyphenylacetic acid, 3′-hydroxyphenylacetic acid, 3′,4′-dihydroxyphenylacetic acid, 3-phenylpropanoic acid, 3-(4′-hydroxyphenyl)propanoic acid, 3-(3′-hydroxyphenyl)propanoic acid, 3-(3′,4′-dihydroxyphenyl)propanoic acid (aka dihydrocaffeic acid), 4-hydroxybenzoic acid, 3-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid (aka protocatechuic acid), and benzene-1,3,5-triol (aka phloroglucinol) were obtained from Sigma-Aldrich (St Louis, MO, USA). Isorhamnetin was from PhytoLab GmbH (Vestenbergsgreuth, Germany). All solvents and water were UHPLC-grade and were purchased from VWR International (Milan, Italy).
8
Bifidobacterium adolescentis Strains Characterization
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Two B. adolescentis strains were used in this work. The strain B. adolescentis IPLA60004, from here onward IPLA60004, belongs to IPLA-CSIC culture collection. IPLA60004 was previously isolated from a human colonic biopsy and was demonstrated to have a high capability to convert the GABA precursor monosodium glutamate (MSG) into GABA (19 (link), 29 (link)). The strain B. adolescentis LMG10502T, from here onward LMG10502T, was purchased from the Belgium culture collection (BCCM/LMG Bacteria Collection, Belgium) and was previously demonstrated to lack gad genes, and hence, the capacity to produce GABA. The strains were routinely grown in MRSc medium [MRS (Biokar, France) supplemented with 0.25% (wt/vol) L-cysteine hydrochloride monohydrate (Sigma-Merck, Germany)]. Stocks stored at −80°C were resuscitated in agar-MRSc in anaerobic jar with anaerocult A (Merck, Germany) at 37°C for 2 days. Isolated colonies were inoculated in 50 mL of MRSc broth and incubated under the same conditions for 20 ± 1 h to obtain the bifidobacterial cultures.
9
Isolation and Cryopreservation of Anaerobic Bacteria
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Depending on the number of available faecal pellets, samples were re-suspended in either 450 or 900 μl of sterile phosphate buffer saline (Sigma-Aldrich, UK) and used to produce tenfold serial dilutions (neat - 10-4). The samples were then vortexed for 30 s and mixed using on a shaker at 1600 rpm. An aliquot of each dilution (100 μl) was plated onto Brain Heart Infusion (BHI) (Oxoid, UK) agar supplemented with mupirocin (50 mg/l) (Sigma-Aldrich, UK), l -cysteine hydrochloride monohydrate (50 mg/l) (Sigma-Aldrich, UK) and sodium iodoacetate (7.5 mg/l) (Sigma-Aldrich, UK) and incubated in an anaerobic cabinet for 48–72 h (Baker Ruskinn, UK). Three colonies from each dilution were randomly selected and streaked to purity on BHI agar supplemented with l -cysteine hydrochloride monohydrate (50 mg/l). Pure cultures were stored in cryogenic tubes at −80 °C.
10
Peptide Modification and Purification
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Example 2
Alternatively, peptides were purified using HPLC and following isolation they were modified with a molecular scaffold reagent containing Michael acceptors. For this, linear peptide was diluted with 50:50 MeCN:H2O up to ˜35 mL, ˜500 μL of 100 mM molecular scaffold reagent containing Michael acceptors in acetonitrile was added, and the reaction was initiated with 5 mL of 1 M NH4HCO3 in H2O. The reaction was allowed to proceed for ˜30-60 min at RT, and lyophilized once the reaction had completed (as judged by MALDI). Once completed, 1 mL of 1M L-Cysteine hydrochloride monohydrate (Sigma) in H2O was added to the reaction for ˜60 min at RT to quench any excess molecular scaffold reagent containing Michael acceptors.
Following lyophilization, the modified peptide was purified as above, while replacing the Luna C8 with a Gemini C18 column (Phenomenex), and changing the acid to 0.1% trifluoroacetic acid. Pure fractions containing the correct desired product were pooled, lyophilized and kept at −20° C. for storage.
All amino acids, unless noted otherwise, were used in the L-configurations.
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