E. coli Nissle 1917 (EcN) was purchased from Ardeypharm GmbH and made chemically competent through treatment with Magnesium and Calcium Chloride (Sigma) following a previously published protocol [32 ]. EcN carrying the LldGFP plasmid was grown to mid-log phase in LB media supplemented with 100 μg/ml Carbenicillin at 37°C with shaking. EcN control without sensing plasmid was grown in LB without carbenicillin. When the culture had an OD600 of 0.55 it was spun down at 4,000g for 5 mins and resuspended in the same volume of artificial saliva with mucin (Pickering Laboratories, 1700–0316) with various concentrations of sodium lactate (Sigma 71718). The artificial saliva had a mineral composition and pH similar to human saliva, with mucin added to increase viscosity. Cells were incubated at 37°C with shaking to allow them to sense the lactate.
Magnesium and calcium chloride
Manufactured by Merck Group
Sourced in Denmark
Magnesium and Calcium Chloride are inorganic compounds commonly used as laboratory reagents. Magnesium Chloride is a white, crystalline solid with the chemical formula MgCl2. Calcium Chloride is a colorless, crystalline solid with the chemical formula CaCl2. Both compounds are highly soluble in water and are used in various applications, including as desiccants, electrolytes, and in the preparation of other chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Rolipram, by Merck Group (1 mentions)
Fenoterol, by Merck Group (1 mentions)
Alprenolol, by Merck Group (1 mentions)
Zaprinast, by Merck Group (1 mentions)
3h dihydroalprenolol, by PerkinElmer (1 mentions)
Salmeterol, by Merck Group (1 mentions)
Ritodrine, by Merck Group (1 mentions)
Geneticin g418, by Thermo Fisher Scientific (1 mentions)
Glp 1 7 36, by Merck Group (1 mentions)
3 isobutyl 1 methyl xanthine (ibmx), by Merck Group (1 mentions)
Envision xcite multilabel plate reader, by PerkinElmer (1 mentions)
Hepes, by Merck Group (1 mentions)
Zeocin, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Caffeine, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Glucagon, by Merck Group (1 mentions)
Black 384 well plate, by Greiner (1 mentions)
2 protocols using magnesium and calcium chloride
1
E. coli Lactate Sensing in Artificial Saliva
2
β-Adrenoceptor Activation Assay
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Isoproterenol, forskolin, ICI 118-551 (ICI), epinephrine, norepinephrine, salbutamol, salmeterol, ritodrine, fenoterol, alprenolol, glucagon, oxyntomodulin, GLP-1-(7-36), rolipram, zaprinast, caffeine, HEPES, IBMX, magnesium and calcium chloride, bovine serum albumin (BSA), and Whatman glass microfiber filters (grade GF/B) were purchased from Sigma-Aldrich (Broendby, Denmark). Dulbecco's modified Eagle's medium (DMEM), Hank's balanced salt solution (HBSS), fetal bovine serum (FBS), zeocin, geneticin G418, and penicillin/streptomycin were purchased from Invitrogen (Naerum, Denmark). Black 384-well plates were purchased from Greiner Bio-One (Kremsmünster, Austria). [ 3 H] dihydroalprenolol and EnVision Xcite multilabel plate reader were from PerkinElmer (Waltham, MA).
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