All bacterial strains are listed in Table 1 . All strains for growth analysis were derivatives of Salmonella enterica subsp. enterica serovar Typhimurium LT2 (hereafter S. enterica). S. enterica strains were grown at 37°C on lysogeny broth (LB, Difco) or no-carbon essential (NCE) minimal medium (Berkowitz et al., 1968 (link)) supplemented with glycerol (22 mM), MgSO4 (concentrations varied and noted in the figure legends), and Wolfe’s trace minerals (1x) (Atlas, 1995 ). Escherichia coli C41 (λDE3) (Miroux and Walker, 1996 (link)) was used for protein overexpression and E. coli DH5α (New England Biolabs) was used for plasmid construction. All E. coli strains were grown at 37°C in LB. Antibiotics for all media were used at the following concentrations: ampicillin, 100 mg mL-1; chloramphenicol, 20 mg mL-1. All chemicals were purchased from Sigma-Aldrich unless otherwise noted; isopropyl β-D -1-thiogalactopyranpside (IPTG, Gold BioTechnology), glycerol (Fisher Scientific), [1-14C] acetyl-CoA (Moravek, Inc.), HEPES (Gold BioTechnology). OAS and L -CysHCl solutions were made fresh in HEPES (pH 7 at 25°C) immediately before each in vitro or in vivo experiment.
Hepes
Manufactured by GoldBio
Sourced in United States
HEPES is a widely used buffering agent in biological research. It is a zwitterionic organic chemical compound that helps maintain a stable pH in aqueous solutions, typically within a range of 6.8 to 8.2. HEPES is commonly used in cell culture media, biochemical assays, and various other laboratory applications that require a well-controlled pH environment.
Automatically generated - may contain errors
Lab products found in correlation
Glycerol, by Thermo Fisher Scientific (1 mentions)
E coli dh5α, by New England Biolabs (1 mentions)
Kanamycin, by GoldBio (1 mentions)
Protease inhibitor cocktail, by Abcam (1 mentions)
Imidazole, by Merck Group (1 mentions)
Bodipy fl gtp, by Thermo Fisher Scientific (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by GoldBio (1 mentions)
Deuterium oxide, by Cambridge Isotopes (1 mentions)
Lysozyme, by GoldBio (1 mentions)
Bodipy fl gtpγs, by Thermo Fisher Scientific (1 mentions)
Leupeptin, by GoldBio (1 mentions)
Mgcl2, by Merck Group (1 mentions)
Acetonitrile, by Avantor (1 mentions)
Formic acid, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Phosphoric acid, by Merck Group (1 mentions)
Benzamidine, by Merck Group (1 mentions)
Dnase 1, by Roche (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Duchefa Biochemie (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Thermo Fisher Scientific (1 mentions)
3 protocols using hepes
1
Salmonella enterica Growth Analysis
2
Purification of Recombinant Proteins
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The protease inhibitor cocktail was purchased from BioVision (Mountain View, CA, USA). Deuterium oxide was purchased from Cambridge Isotope Laboratories (Tewksbury, MA, USA). Chelating sepharose resin for Ni-IDA purification was purchased from Cytiva (Uppsala, Sweden). EDTA, Tris, and NaH2PO4 were purchased from Duchefa Biochemie BV (Haarlem, The Netherlands). HEPES, IPTG, leupeptin, lysozyme, kanamycin, and TCEP were purchased from Goldbio (St. Louis, MO, USA). Acetonitrile, H2O, and MeOH were purchased from JT Baker (Phillipsburg, NJ, USA). DNase I was purchased from Roche (Basel, Switzerland). BODIPY-FL-GTP and BODIPY-FL-GTPγS were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Acetic acid, benzamidine, formic acid, GDP, phosphoric acid, imidazole, and MgCl2 were purchased from Sigma–Aldrich (St. Louis, MO, USA).
3
Reconstitution of Microtubule Dynamics
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GTP and paclitaxel (Taxol) were purchased from Cytoskeleton (Denver, CO). Guanosine-5′-[(α,β)-methyleno]triphosphate, sodium salt (GMPCPP), the nonhydrolyzable analog of GTP, was purchased from Jena Bioscience (Jena, Germany). Piperazine-N, N′-bis (2-ethanesulfonic acid) (PIPES), ATP, and glucose were purchased from Sigma (St. Louis, MO). HEPES and dithiothreitol (DTT) were purchased from GoldBio (St Louis, MO). EGTA, MgCl2, and KCl were purchased from Thermo Fisher Scientific (Waltham, MA). Sterile collodion (nitrocellulose) 2% in amyl acetate and amyl acetate were purchased from Electron Microscopy Sciences (Hatfield, PA). d-Biotin was purchased from Avidity (Aurora, CO). Chloroform solutions of 1,2-dioleoyl-sn-glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl), mini extruder, Nuclepore track-etch membrane (50 nm) from Whatman, and filter support disks 10 mm from Whatman were purchased from Avanti Polar Lipids (Alabaster, AL). High vacuum grease, silicone elastomer base (184 Sylgard), and the corresponding elastomer curing agent were manufactured by Dow Corning (Midland, MI). 2% Dimethyldichlorosilane in octamethylcyclooctasilane (PlusOne Repel-Silane ES) was purchased by GE Healthcare (Chicago, IL). Optical adhesive Norland 65 was purchased from Norland Products (Cranbury, NJ).
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