S. aureus (ATCC26112) was obtained from the Chinese Medicine Bacterial Preservation Centre (Beijing, China). Fraxetin, at 99% purity, was purchased from Nuowei Xin (Dalian, China). The fraxetin was dissolved in absolute ethanol and concentrated solutions were added to the bacterial cultures to maintain the lowest possible concentration of ethanol in the cultures. The restriction enzyme, pBR322, was purchased from Takara Bio, Inc. (Shiga, Japan). DAPI was purchased from Beyotime Institute of Biotechnology (Shanghai, China). Common chemicals (ethanol, NaCl, KCl, KH2PO4, K2HPO4, beef extract, peptone) were purchased from Tianjin Kemiou Chemical Reagent Co, Ltd. (Tianjin, China). SDS, Tris-base, bovine serum albumin, adenosine triphosphate and proteinase K were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). All assays were performed according to the manufacturer’s instructions.
Adenosine triphosphate (atp)
Manufactured by Sangon
Sourced in China
ATP (Adenosine Triphosphate) is a molecule that serves as the primary energy currency in living cells. It is essential for various cellular processes, including metabolism, signaling, and transport. ATP is a nucleotide composed of adenosine and three phosphate groups, and its hydrolysis releases a significant amount of energy, which can be used to power various cellular activities.
Automatically generated - may contain errors
Lab products found in correlation
Mgcl2, by Sangon (3 mentions)
Adenosine diphosphate (adp), by Sangon (3 mentions)
Sodium chloride (nacl), by Sangon (2 mentions)
Milli q, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Sangon (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Potassium chloride (kcl), by Tianjin Kemiou Chemical Reagent (1 mentions)
Pbr322, by Takara Bio (1 mentions)
Sodium dodecyl sulfate (sds), by Sangon (1 mentions)
Sodium chloride (nacl), by Tianjin Kemiou Chemical Reagent (1 mentions)
Ethanol, by Tianjin Kemiou Chemical Reagent (1 mentions)
Proteinase k, by Sangon (1 mentions)
Kh2po4, by Tianjin Kemiou Chemical Reagent (1 mentions)
K2hpo4, by Tianjin Kemiou Chemical Reagent (1 mentions)
Tris base, by Sangon (1 mentions)
Methyl 3h sam, by PerkinElmer (1 mentions)
Superdex 200 column, by GE Healthcare (1 mentions)
Cytidine triphosphate, by Sangon (1 mentions)
Dntp mixture, by Takara Bio (1 mentions)
Pet22b vector, by Merck Group (1 mentions)
16 protocols using adenosine triphosphate (atp)
1
Antibacterial activity of fraxetin against S. aureus
2
Optimizing Crystallization Conditions for Methyltransferase Enzyme
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SAM, SAH, 5′-GMP, Tris-base, β-mercaptoethanol (β-Me), KCl and the reagents used to optimize crystallization conditions were purchased from Sigma–Aldrich (St Louis, MO, USA). MgCl2, NaCl, adenosine triphosphate, cytidine triphosphate, guanosine triphosphate and uridine triphosphate were from Sangon Biotech (Shanghai, China). [Methyl-3H] SAM was obtained from PerkinElmer (Waltham, MA, USA); crystallization kits were from Hampton research (Aliso Viejo, CA, USA). Primers for polymerase chain reaction were synthesized by Invitrogen (Shanghai, China) and Biosune (Shanghai, China); the pET22b vector was from Merck–Millipore (Darmstadt, Germany). The KOD-plus mutagenesis kit, Pyrobest DNA polymerase and dNTP mixture were obtained from Takara (Shiga, Japan); T4 ligase and other restriction endonucleases were obtained from MBI Fermentas (Pittsburgh, PA, USA). Pyrophosphatase (PPiase) was purchased from Roche Applied Science (Basel, Switzerland). Ni2+-NTA Superflow was obtained from Qiagen (Dusseldorf, Germany). The Superdex 200 column and 3 mm filter papers were from GE Healthcare (Fairfield, CT, USA). The analog of SAM, sinefungin, was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
3
Oligonucleotide Purification and Characterization
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ATP, GTP, CTP, UTP and all the oligonucleotides were purchased from Shanghai Sangon Biotechnology Co. Ltd. (China). The oligonucleotides were purified using high performance liquid chromatography and their sequences are listed in Table S1 in the ESI.† All other reagents were of analytical grade and used without further purification. Ultrapure water obtained from a Millipore water purification system (≥18 MΩ, Milli-Q, Millipore) was used in all assays. Phosphate buffer saline (PBS, 10 mM pH 7.4) containing 0.1 M NaCl was prepared by mixing the stock solutions of NaH2PO4 and Na2HPO4.
4
Synthesis of Gold Nanoparticles
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Chloroauric acid (HAuCl4·4H2O) was obtained from the Shanghai Chemical Reagent Company (Shanghai, China). Trisodium citrate was provided by the Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). The DNA sequences, ATP, GTP, UTP, CTP, and phosphate-buffered saline (PBS) were purified by Sangon Biotechnology Co., Ltd. (Shanghai, China). All other reagents were of analytical grade, and the aqueous solutions that were used were prepared with ultrapure water (18 MΩ, Milli-Q, Millipore, Shanghai China).
5
Cell Culture and Reagent Conditions
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HEK293T, H1299, and A549 cell lines were obtained from the American Type Culture Collection, and they had been verified. HEK293T cell line was cultured in Dulbecco's modified Eagle's medium (Gibco; C11995500BT) supplemented with 10% fetal bovine serum (FBS) (Hyclone; SV30087.03). A549 cell line was cultured in Dulbecco's modified Eagle's medium/F12 = 1:1 (Gibco; C11330500BT) supplemented with 10% FBS. H1299 cell line was cultured in RPMI1640 (Gibco; C11875500BT) supplemented with 10% FBS. All cells were cultured at 37 °C with 5% carbon dioxide.
Reagents: IPTG (Sangon Biotech; A100487-0005), Kanamycin (Beyotime; A506636-0025), MG132 (MCE; HY-13259), TM (Beyotime; SC0393-10 mM), ATP (Sangon Biotech; A600020), GSK2606414 (MCE; HY-18072), and CHX (MCE; HY-12320).
Reagents: IPTG (Sangon Biotech; A100487-0005), Kanamycin (Beyotime; A506636-0025), MG132 (MCE; HY-13259), TM (Beyotime; SC0393-10 mM), ATP (Sangon Biotech; A600020), GSK2606414 (MCE; HY-18072), and CHX (MCE; HY-12320).
6
ATPase Activity Assay of Purified Proteins
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All ATPase assays were performed using the ATPase/GTPase Activity Assay Kit (Sigma-Aldrich, catalog number MAK113-1KT). Purified SugABC (0.72 μg) or 5.46-μg LpqY-SugABC-E164Q in A8-35 was incubated in a 20-μl reaction volume containing 10 mM Hepes, (pH 7.5), 75 mM NaCl, 1 mM ATP, and 2.5 mM MgCl2 (Sangon Biotech) for 30 min at 37°C. The reaction was stopped by adding 100 μl of reagent (catalog number MAK113A) and incubated for an additional 30 min at room temperature to generate the colorimetric product. Absorbance at 620 nm was measured at room temperature using a SpectraMax iD3 multifunction reader (Molecular Devices). ATPase activity was represented as phosphate (nanomoles) produced by 1 mg of protein per minute. Experiments were performed in triplicate.
7
Graphene Oxide-Assisted Enzymatic Assay
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Nb.bpu10I endonuclease and Bsm DNA polymerase were obtained from Thermo Scientific (Waltham, MA, USA). Graphene oxide (GO) was obtained from Aladdin Reagent Co., Ltd. (Shanghai, China). Deoxyribonucleoside triphosphate (dNTP), ATP, ADP, AMP, guanosine triphosphate (GTP), uridine triphosphate (UTP) and cytidine triphosphate (CTP) were all purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Human serum was purchased from XinFan Bio-technology Co., Ltd. (Beijing, China). In all experimental procedures, ultrapure water (18.2 MΩ cm) acquired from a Micropore filtration system was employed to prepare all solutions.
In this work, all the DNA sequences were synthesized and modified by Sangon Biotech Co., Ltd. (Shanghai, China). In theSupplementary Information , these sequences are presented in Table S1 .
In this work, all the DNA sequences were synthesized and modified by Sangon Biotech Co., Ltd. (Shanghai, China). In the
8
GO-Based Enzyme Biosensing Platform
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Graphene oxide (GO) was obtained from Shanghai Tanyuanhuigu Co., Ltd. (Shanghai, China). Exonuclease III (Exo III) and cAMP-dependent protein kinase (PKA) were obtained from New England Biolabs Inc. (Ipswich, MA, United States). Cysteine-terminated kemptide (LRRASLGGGGC) and ATP were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Thrombin, hemoglobin (Hb), bovine serum albumin (BSA) and fetal calf serum (FBS) were supplied by Dingguo Biotech Co., Ltd. (Beijing, China). Dopamine hydrochloride and N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride (H-89) were purchased from Aladdin Reagents Inc. (Shanghai, China). Tris-(hydroxymethyl)aminomethane (Tris), AgNO3 and Ti(SO4)2 were supplied by Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Ultrapure water was always used and obtained from a Milli-Q water purification system (Millipore Corp., Bedford, MA, United States).
9
Cultivation and Expression of E. coli Rosetta (DE3)
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Escherichia coli cultivation medium (LB) was purchased from Oxoid (The UK). E. coli Rosetta (DE3) and plasmid pET-28a were both
purchased from Novagen (The USA). Isopropy1 β-d -thiogalactopyranoside
(IPTG) was purchased from Shanghai Jinsui Biotechnology Co., Ltd.
(Shanghai, China). ATP, AMP, NAD+, kanamycin, chloromycetin,
sodium hexametaphosphate (polyP6), Tris, and glucose-6-phosphate
dehydrogenase were purchased from Sangon Biotech Co., Ltd. (Shanghai,
China). All other chemicals were of analytical grade from commercial
sources and used without further purification.
purchased from Novagen (The USA). Isopropy1 β-
(IPTG) was purchased from Shanghai Jinsui Biotechnology Co., Ltd.
(Shanghai, China). ATP, AMP, NAD+, kanamycin, chloromycetin,
sodium hexametaphosphate (polyP6), Tris, and glucose-6-phosphate
dehydrogenase were purchased from Sangon Biotech Co., Ltd. (Shanghai,
China). All other chemicals were of analytical grade from commercial
sources and used without further purification.
10
Cellular Inflammation and Apoptosis Assay
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Lipopolysaccharide, 2,7-dichlorofluorescin diacetate, N-acetylcysteine, rabbit antibody against human GSDMD (G7422); FITC-conjugated goat antibody against rabbit IgG (F9887); and Cy3-conjugated sheep antibody against mouse IgG (C2181) were obtained from Sigma-Aldrich. ATP, propidium iodide, and colchicine were purchased from Sangon Biotech. Nigericin and poly(dA:dT) were obtained from InvivoGen. MitoTracker Deep Red, MitoTracker Green, MitoTracker Red CMXRos, Hochest 33342, Lipofectamine 2000, Lipofectamine RNAiMax, TMRM, and Carboxy-H2DCFDA were obtained from Invitrogen. ViaFect transfection reagent was purchased from Promega. The antibody against IL-1β (5129) was purchased from BioVision. Antibodies against Nlrp3 (ab4207) and Hsp60 (ab46798), Alexa Fluor 594-conjugated donkey antibody to rabbit IgG (ab150076), and the antibody against COX IV (ab33985) were obtained from Abcam. The antibody against caspase-1 (5B10) was purchased from eBioscience. The antibody against β-actin (I102) was purchased from Bioworld Technology. The antibody against LC3 (NB100-2220) was obtained from Novus. The antibody against NLRP3 (MAB7578) was obtained from R&D Systems. M-CSF was purchased from Peprotech. Alexa Fluor 633-conjugated donkey antibody to goat IgG (705-175-003) was purchased from Jackson ImmunoResearch. 3-MA and linezolid were obtained from Selleck.
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