For the transport and preparation of the smooth muscle strips and for the incubation in the organ bath, KH was used (in mmol/L: NaCl 115.0 (VWR International GmbH, Darmstadt, Germany), 4.7 KCl, 2.5 CaCl2, 1.2 MgCl2, 1.2 NaH2PO4, 25.0 NaHCO3 (all Carl Roth GmbH & Co. KG, Karlsruhe, Germany) and 11.0 glucose (VWR International GmbH, Darmstadt, Germany)). Before use, the buffer was gassed with carbogen (95% O2, 5% CO2) under constant temperature conditions (38 °C). The final pH of the buffer was 7.4. Furthermore, two buffer variations were used for the experiments: a calcium-free buffer (in mmol/L: NaCl 115.0 (VWR International GmbH, Darmstadt, Germany), 4.7 KCl, 1.2 MgCl2, 1.2 NaH2PO4, 25.0 NaHCO3 (all Carl Roth GmbH & Co. KG, Karlsruhe, Germany) and 11.0 glucose (VWR International GmbH, Darmstadt, Germany)) and a potassium-enriched buffer, containing potassium chloride instead of sodium chloride (in mmol/L: 123.4 KCl, 2.5 CaCl2, 1.2 MgCl2, 1.2 NaH2PO4, 25.0 NaHCO3 (all Carl Roth GmbH & Co. KG, Karlsruhe, Germany) and 11.0 glucose (VWR International GmbH, Darmstadt, Germany)).
Mgcl2
Manufactured by Carl Roth
Sourced in Germany, United States
Magnesium chloride (MgCl2) is an inorganic compound with the chemical formula MgCl2. It is a source of magnesium ions (Mg2+) and chloride ions (Cl-) and is commonly used in various laboratory and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Carl Roth (12 mentions)
Hepes, by Carl Roth (10 mentions)
Cacl2, by Carl Roth (10 mentions)
Potassium chloride (kcl), by Carl Roth (7 mentions)
Nahco3, by Carl Roth (4 mentions)
Cacl2, by Merck Group (4 mentions)
Hepes, by Merck Group (4 mentions)
Sucrose, by Carl Roth (3 mentions)
Glucose, by Carl Roth (3 mentions)
Nah2po4, by Carl Roth (2 mentions)
K2hpo4, by Carl Roth (2 mentions)
Mannitol, by Carl Roth (2 mentions)
Glutamic acid, by Merck Group (2 mentions)
Amytal, by Merck Group (2 mentions)
Malic acid, by Merck Group (2 mentions)
Succinic acid, by Merck Group (2 mentions)
Adenosine 5 diphosphate sodium salt, by Merck Group (2 mentions)
Kh2po4, by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
Formaldehyde, by Carl Roth (2 mentions)
33 protocols using mgcl2
1
Smooth Muscle Contractility Assay Using Krebs-Henseleit Buffer
2
Preparation of MXene Suspensions
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3
Integrin Ligand-Mediated Virus Binding Inhibition
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For the binding inhibition assay, cells were seeded on 24-well plates with DMEM supplemented with 2% FBS at a concentration of 1 × 105 cells per well, 12–16 h prior to virus inoculation. Medium was replaced by serum free DMEM medium and cells were pre-incubated at 4 °C for 15 min prior to treatment with integrin ligands. Type I collagen (0–500 μg/mL-Sigma-Aldrich, St. Louis, MO, USA), synthetic Arg-Gly-Asp (RGD motif) tripeptide (0–250 μg/mL-Sigma-Aldrich, St. Louis, MO, USA) and recombinant mouse vitronectin (0–50 μg/mL-Sino Biological, Beijing, China) were added into the wells with serum free DMEM supplemented with 1 mM MnCl2 (Carl-Roth, Karlsruhe, Germany) and 1 mM MgCl2 (Carl-Roth, Karlsruhe, Germany) and incubated at 4 °C for 30 min to allow ligand binding. After this, plates were placed on ice and washed twice, and cells were inoculated with flaviviruses at a MOI of 10. Cells were incubated for 1 h on ice with constant agitation every 15 min. After this period, the inoculum was removed and the wells were washed four times with cold 1× PBS to remove unbound virus particles. Cell monolayers were resuspended in RLT buffer (Qiagen, Hilden, Germany) and stored at −80 °C.
4
Kinetic Characterization of Rab Prenylation
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200 nM RABGGT was incubated with 200 nM CHM and 800 nM {3,7,11-trimethyl-12-(7-nitro-benzo[1,2,5]oxadiazo-4-ylamino),-dodeca-2,6,10-trien-1} pyrophosphate (NBP-FPP) (Jena Bioscience, LI-013) in prenylation buffer containing 20 mM HEPES (Carl Roth, 6763.3), pH 7.2, 25 mM NaCl (Carl Roth, P029.2), 2 mM DTE (Sigma, D8255), 2 mM MgCl2 (Carl Roth, HN03.1) for 5 min. The prenylation was initiated by adding 250 nM RAB33B [53 (link)]. The fluorescence signal upon the incorporation of {3,7,11-trimethyl-12-(7-nitro-benzo[1,2,5]oxadiazo-4-ylamino),-dodeca-2,6,10-trien-1} pyrophosphate (NBP-FPP) into RAB33B was measured on a Spex Fluoromax-3 spectrofluorometer (Jobin Yvon, Edison). Excitation and emission monochromators were set to 480 nm and 520 nm, respectively. The progress curves were fit to single exponential function using Prism version 7.00 for Windows in order to obtain the half-life of a reaction (t1/2).
5
Lentiviral Vector Concentration and Purification
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The lentiviral vector was harvested 72 h post-transfection. DENARASE® (c-Lecta) and MgCl2 (Carl Roth) were added to the cell culture broth one hour before harvest at a final concentration of 10 U/mL and 2 mM, respectively. After nucleic acid digestion, the lentiviral vector containing cell culture broth was directly clarified with Sartoclear Dynamics® Lab V50 (0.45 μm polyethersulfone membrane version) with 5 g/L diatomaceous earth (Sartorius). The lentiviral vector was aliquoted and stored at -80°C.
6
Haloferax volcanii Cultivation Protocol
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Haloferax volcanii strain H26 was kindly provided by Thorsten Allers (University of Nottingham, UK). It was and grown in complex medium [36] (link) or in synthetic medium [37] (link) supplemented with 8 µM FeSO4 (Roth, P015.1), 0,1% (v/v) SL-6 trace element solution [38] (link) (all from Roth), 1 ml vitamin solution (Sigma Aldrich, B6891), 50 µg ml−1 uracil (Applichem, A0667) and 100 mM MOPS pH 7.2 (Sigma Aldrich, M3183). All components of the synthetic medium were of the grade “per analysis” and thus free of phosphate, e.g. K2HPO4 (Roth, 6878.2), NH4Cl (Applichem, A0988), glucose (Merck, 1083441000), NaCl (Roth, 3957.5), MgCl2 (Roth, 2189.1), MgSO4 (Applichem, A1037), KCl (Roth, 6781,1), CaCl2 (Applichem, A3587), and Tris (A1086). If not otherwise stated, the synthetic medium was also supplemented with 0.5% (w/v) glucose as a C source, 10 mM NH4Cl as a N source, and 1 mM K2HPO4 as a P source. For growth experiments with DNA as a source of P K2HPO4 was omitted and genomic DNA was added to a final concentration of 250 µg/ml. Cultures were grown in Erlenmeyer flasks in a rotary shaker at 42 °C and 250 rpm or in microtiter plates as described below.
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Haloferax volcanii strain H26 was kindly provided by Thorsten Allers (University of Nottingham, UK). It was and grown in complex medium [36] (link) or in synthetic medium [37] (link) supplemented with 8 µM FeSO4 (Roth, P015.1), 0,1% (v/v) SL-6 trace element solution [38] (link) (all from Roth), 1 ml vitamin solution (Sigma Aldrich, B6891), 50 µg ml−1 uracil (Applichem, A0667) and 100 mM MOPS pH 7.2 (Sigma Aldrich, M3183). All components of the synthetic medium were of the grade “per analysis” and thus free of phosphate, e.g. K2HPO4 (Roth, 6878.2), NH4Cl (Applichem, A0988), glucose (Merck, 1083441000), NaCl (Roth, 3957.5), MgCl2 (Roth, 2189.1), MgSO4 (Applichem, A1037), KCl (Roth, 6781,1), CaCl2 (Applichem, A3587), and Tris (A1086). If not otherwise stated, the synthetic medium was also supplemented with 0.5% (w/v) glucose as a C source, 10 mM NH4Cl as a N source, and 1 mM K2HPO4 as a P source. For growth experiments with DNA as a source of P K2HPO4 was omitted and genomic DNA was added to a final concentration of 250 µg/ml. Cultures were grown in Erlenmeyer flasks in a rotary shaker at 42 °C and 250 rpm or in microtiter plates as described below.
7
Fluorescence Substrate Preparation Protocol
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The fluorescence susbstrates were purchased in DNA-synthesis, Moscow, Russia. Other oligonucleotides were purchased in Evrogen, Moscow, Russia. DNAse/RNAse free water was purchased from Qiagen, Hilden, Germany. Salts, including MgCl2, NaCl, KCl, HEPES, and DEPC were purchased from CarlROTH, Carlsruhe, Germany. Acrylamide was purchased from Applichem, Illinois, USA. Other materials for electrophoresis, including agarose, tris, boric acid, EDTA, bisAcrylamide, APS, TEMED, ethidium bromide, and SDS were purchased from Helicon, Moscow, Russia. Ladders and gel loading dyes were purchased from Evrogen, Moscow, Russia. Consumables for bacteria growth, including agar, tripton and yeast extract, were purchased in Dia-M, Moscow, Russia. Glass beads of technical grade was a gift from a technical department. Sodium acetate, phenol, and chlorophorm were purchased from Lenreactiv, Saint-Petersburg, Russia.
8
Chemical Reagents Sourcing Protocol
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All chemicals were obtained from Sigma-Aldrich (Merck, Darmstadt, Germany) except PIPES ((Piperazine-N,N’-bis-(2-ethanesulphonic acid)), HEPES (N-2-Hydroxyethyl piperazine-N’-2-ethane sulphonic acid), EGTA (ethylene glycol bis(2-aminoethyl ether)-N,N,N’,N’-tetraacetic acid) and MgCl2 which were received from Carl Roth (Karlsruhe, Germany).
9
Mitochondrial Bioenergetics and Antioxidant Analysis
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Glutamic acid, succinic acid, adenosine-5′-diphosphate sodium salt (ADP), cytochrome c from bovine heart, malic acid, ethylene glycol-bis-(b-aminoethylether)-N,N,N′N′-tetra acetic acid (EGTA), KH2PO4, TrisHCl, ethylenediamine tetra-acetic acid (EDTA), amytal, carboxyatractyloside (CAT), guanosine triphosphate (GTP)quercetin, acetonitrile, 2,2-azinobis (ethyl-2,3-dihydrobenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), potassium persulfate, iron(III) chloride hexahydrate (FeCl3·6H2O), 2,4,6-tripyridyl-s-triazine (TPTZ), hydrochloric acid, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) were obtained from “Sigma–Aldrich GmbH”, Steinheim, Germany. Sucrose, mannitol, HEPES, KCl, MgCl2, trifluoroacetic acid, hyperoside, isoquercitrin, rutin were obtained from “Carl Roth Gmbh”, Karlsruhe, Germany.
10
Hematopoietic Growth Factor Protocol
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Recombinant human SCF, IL-3 and EPO were from Peprotech (Hamburg, Germany). Insulin solution, holo-transferrin, heparin, hydrocortisone, bovine albumin, crystal violet, and DEAE-Dextran were from Sigma/Aldrich (Schnelldorf, Germany). MgCl2, CaCl2, NaHCO3, and formaldehyde were from Carl Roth (Karlsruhe, Germany).
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