Structured Review

Thermo Fisher zncl2
Zncl2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 15 article reviews
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zncl2 - by Bioz Stars, 2019-10
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Related Articles

MMP-2 Human Biotrak Assay:

Article Title: Inhibitory effect of norcantharidin on melanoma tumor growth and vasculogenic mimicry by suppressing MMP-2 expression
Article Snippet: The gel was washed twice in 2.5% (w/v) Triton X-100 solution and incubated overnight at 37°C in developing buffer [50 mmol/l Tris/HCl (pH 7.4), 10 mmol/l CaCl2 , 5 mmol/l ZnCl2 and 0.05% Brij™ 35 Surfact-Amps™ Detergent Solution (Thermo Fisher Scientific, Inc.)]. .. The gels were subsequently stained with Coomassie Brilliant Blue R250 and destained until the wash remained clear and cleared zones associated with MMP activity were apparent.

Centrifugation:

Article Title: A Re‐evaluation of Electron‐Transfer Mechanisms in Microbial Electrochemistry: Shewanella Releases Iron that Mediates Extracellular Electron Transfer
Article Snippet: The cells were pelleted by centrifugation (3220 g, 15 min) and washed twice with MOPS buffer (10 mL; 20 mm MOPS, 30 mm Na2 SO4 , pH 7.4) before re‐suspension in MOPS buffer (5 mL). .. The trace minerals solution contained (per liter): 1 g FeCl2 ⋅4 H2 O, 0.5 g MnCl2 ⋅4 H2 O, 0.3 g CoCl2 ⋅6 H2 O, 0.2 g ZnCl2 , 0.05 g Na2 MoO4 ⋅2 H2 O (ACROS organics), 0.02 g H3 BO3 , 0.1 g NiSO4 ⋅6 H2 O, 0.002 g CuCl2 ⋅2 H2 O (ACROS organics), 0.006 g Na2 SeO3 ⋅5 H2 O and 0.008 g Na2 WO4 ⋅2 H2 O (ACROS organics).

Zymography:

Article Title: Inhibitory effect of norcantharidin on melanoma tumor growth and vasculogenic mimicry by suppressing MMP-2 expression
Article Snippet: Paragraph title: Zymography assays and MMP-2 protein concentration determination ... The gel was washed twice in 2.5% (w/v) Triton X-100 solution and incubated overnight at 37°C in developing buffer [50 mmol/l Tris/HCl (pH 7.4), 10 mmol/l CaCl2 , 5 mmol/l ZnCl2 and 0.05% Brij™ 35 Surfact-Amps™ Detergent Solution (Thermo Fisher Scientific, Inc.)].

Autoradiography:

Article Title: CRISPRscan: designing highly efficient sgRNAs for CRISPR/Cas9 targeting in vivo
Article Snippet: To produce 5′-end-labeled sgRNAs, purified sgRNA transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BIoLabs) to 50 pmol of sgRNA in a final volume of 10 μL containing 50 mM Bis-Propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20U, Invitrogen). .. The reaction was stopped by adding formamide dye buffer (95% formamide, 10 mM EDTA, 0.025% bromophenol blue and 0.025% xylene cyanol), and the radiolabeled sgRNA purified by 8% polyacrylamide gel electrophoresis.

Article Title: New scoring system to identify RNA G-quadruplex folding
Article Snippet: Before radioactive 5′-end-labelling, 50 pmol of purified RNA transcripts were dephosphorylated using 1 U of antarctic phosphatase (New England BioLabs) in a 10-µl final volume reaction containing 50 mM Bis-Tris-propane, pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and 20 U RNase OUT (Invitrogen). .. Before radioactive 5′-end-labelling, 50 pmol of purified RNA transcripts were dephosphorylated using 1 U of antarctic phosphatase (New England BioLabs) in a 10-µl final volume reaction containing 50 mM Bis-Tris-propane, pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and 20 U RNase OUT (Invitrogen).

Article Title: Exploring mRNA 3?-UTR G-quadruplexes: evidence of roles in both alternative polyadenylation and mRNA shortening
Article Snippet: To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen). .. The reactions were stopped by the addition of formamide dye buffer (95% formamide, 10 mM EDTA, 0.025% bromophenol blue and 0.025 xylene cyanol), and the RNA molecules were then purified by 10% polyacrylamide–8 M urea gel electrophoresis.

Article Title: 5?-UTR G-quadruplex structures acting as translational repressors
Article Snippet: In order to produce 5′-end-labeled PG4s, purified transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BioLabs,) to 50 pmol of RNA and incubating the reaction mixture for 30 min at 37°C in a final volume of 10 µl containing 50 mM Bis-Propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen). .. The reactions were stopped by adding formamide dye buffer (95% formamide, 10 mM EDTA, 0.025% bromophenol blue and 0.025% xylene cyanol), and the RNA molecules purified by 10% polyacrylamide gel electrophoresis.

Article Title: 3′ UTR G-quadruplexes regulate miRNA binding
Article Snippet: Purified transcripts (50 pmol) were dephosphorylated for 1 h at 37°C in a solution containing 1 U of Antarctic phosphatase (New England Biolabs), 50 mM bis-propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 , and RNase OUT (20 U, Invitrogen) in a final volume of 10 µL. .. The reactions were stopped by the addition of 20 µL of formamide dye buffer (95% formamide, 10 mM EDTA, and 0.025% bromophenol blue), and the RNA was then purified by 10% polyacrylamide gel electrophoresis.

Article Title: The folding of 5′-UTR human G-quadruplexes possessing a long central loop
Article Snippet: In order to produce 5′-end-labeled RNA molecules, purified transcripts (50 pmol) were dephosphorylated at 37°C for 30 min by adding 5 units of antarctic phosphatase (New England BioLabs) in a final volume of 10 μL containing 50 mM Bis-propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 , and 20 units RNase OUT (Invitrogen). .. The RNAs molecules were purified by 10% polyacrylamide 8 M urea gel electrophoresis.

Construct:

Article Title: Interaction of catechol and non-catechol substrates with externally or internally facing dopamine transporters
Article Snippet: The pCIN4 vector and the pCIN4-synthetic human DAT construct (pCIN4-DAT) were generous gifts from Dr. Jonathan A. Javitch (Columbia University, New York, NY, USA). .. Digitonin and ZnCl2 were from Acros Organics Co. (Geel, Belgium).

Real-time Polymerase Chain Reaction:

Article Title: Engineering glycoside hydrolase stability by the introduction of zinc binding
Article Snippet: The figures showing structural models were created using PyMOL (v.1.7.2.1; Schrödinger). .. DSF assays comparing the thermal stability of J30 wt and J30 CCH were performed on an Applied Biosciences StepOnePlus Real-Time PCR System in triplicate on a 30 µl scale containing 0.5 mg ml−1 enzyme, 150 mM NaCl, 25 mM HEPES pH 7.4, 10 µM ZnCl2 and SYPRO Orange Protein Gel Stain (Life Technologies, catalog No. S-6650) at 5×. .. Fluorescence signals were recorded from 25 to 99°C.

Incubation:

Article Title: Inhibitory effect of norcantharidin on melanoma tumor growth and vasculogenic mimicry by suppressing MMP-2 expression
Article Snippet: Gelatin zymography was used to examine the levels of MMP-2 activity in A375 cells that were either untreated (control group) or treated with 28 µg/ml NTCD for 48 h. The culture media were collected and subjected to 10% SDS-PAGE using 0.01% w/v gelatin. .. The gel was washed twice in 2.5% (w/v) Triton X-100 solution and incubated overnight at 37°C in developing buffer [50 mmol/l Tris/HCl (pH 7.4), 10 mmol/l CaCl2 , 5 mmol/l ZnCl2 and 0.05% Brij™ 35 Surfact-Amps™ Detergent Solution (Thermo Fisher Scientific, Inc.)]. .. The gels were subsequently stained with Coomassie Brilliant Blue R250 and destained until the wash remained clear and cleared zones associated with MMP activity were apparent.

Article Title: Snapshot Hyperspectral Light-Sheet Imaging of Signal Transduction in Live Pancreatic Islets
Article Snippet: Briefly, 1 mM dye in dimethyl sulfoxide was diluted in 1 ml Krebs Ringer Bicarbonate HEPES buffer (KRBH) to a final concentration of 4 μ M. To disperse the hydrophobic dye molecules into solution, the final buffer were sonicated for 10 min. Islets were incubated in this buffer at room temperature for 45 min with gentle rocking. .. ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4.

Article Title: Exploring mRNA 3?-UTR G-quadruplexes: evidence of roles in both alternative polyadenylation and mRNA shortening
Article Snippet: The reactions were incubated for 2 h at 37°C followed by a DNase RQ1 (Promega) treatment at 37°C for 20 min. .. To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen).

Article Title: Engineering glycoside hydrolase stability by the introduction of zinc binding
Article Snippet: DSF assays comparing the thermal stability of J30 wt and J30 CCH were performed on an Applied Biosciences StepOnePlus Real-Time PCR System in triplicate on a 30 µl scale containing 0.5 mg ml−1 enzyme, 150 mM NaCl, 25 mM HEPES pH 7.4, 10 µM ZnCl2 and SYPRO Orange Protein Gel Stain (Life Technologies, catalog No. S-6650) at 5×. .. DSF assays comparing the thermal stability of J30 wt and J30 CCH were performed on an Applied Biosciences StepOnePlus Real-Time PCR System in triplicate on a 30 µl scale containing 0.5 mg ml−1 enzyme, 150 mM NaCl, 25 mM HEPES pH 7.4, 10 µM ZnCl2 and SYPRO Orange Protein Gel Stain (Life Technologies, catalog No. S-6650) at 5×.

Activity Assay:

Article Title: Inhibitory effect of norcantharidin on melanoma tumor growth and vasculogenic mimicry by suppressing MMP-2 expression
Article Snippet: Gelatin zymography was used to examine the levels of MMP-2 activity in A375 cells that were either untreated (control group) or treated with 28 µg/ml NTCD for 48 h. The culture media were collected and subjected to 10% SDS-PAGE using 0.01% w/v gelatin. .. The gel was washed twice in 2.5% (w/v) Triton X-100 solution and incubated overnight at 37°C in developing buffer [50 mmol/l Tris/HCl (pH 7.4), 10 mmol/l CaCl2 , 5 mmol/l ZnCl2 and 0.05% Brij™ 35 Surfact-Amps™ Detergent Solution (Thermo Fisher Scientific, Inc.)].

Solubility:

Article Title: Zinc flexes its muscle: Correcting a novel analysis of calcium for zinc interference uncovers a method to measure zinc
Article Snippet: A 4-mM BAPTA (Sigma-Aldrich) stock solution was prepared in 0.3 N NaHCO3 (Thermo Fisher Scientific) to reach an optimal solubility (50 mg/ml) as suggested by Sigma-Aldrich. .. ZnCl2 (Thermo Fisher Scientific) was prepared as either 10 or 25 mM in aqueous solution.

Transfection:

Article Title: Dual Readout BRET/FRET Sensors for Measuring Intracellular Zinc
Article Snippet: Two days after transfection, cells were released from the wells using trypsin (Thermo Fisher Scientific) and resuspended in 1 mL of preheated (37 °C) live-cell imaging buffer (20 mM HEPES (pH 7.4), 140 mM NaCl, 2.5 mM KCl, 1.8 mM CaCl2 , and 1.0 mM MgCl2 ). .. The measurement was performed for 50 min, during which 50 μM N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN, Sigma-Aldrich) was loaded onto the cells after 15 min and 100 μM ZnCl2 and 5 μM 2-mercaptopyridine N-oxide (pyrithione, Acros Organics) after 35 min. During the additions of reagents, the measurement was stopped for ∼1 min. Sensor occupancies were calculated using , and corresponding Zn2+ levels were calculated using , in which R min and R max are the steady-state emission ratios after the addition of TPEN and Zn2+ 2 3

Cell Culture:

Article Title: Inhibitory effect of norcantharidin on melanoma tumor growth and vasculogenic mimicry by suppressing MMP-2 expression
Article Snippet: The gel was washed twice in 2.5% (w/v) Triton X-100 solution and incubated overnight at 37°C in developing buffer [50 mmol/l Tris/HCl (pH 7.4), 10 mmol/l CaCl2 , 5 mmol/l ZnCl2 and 0.05% Brij™ 35 Surfact-Amps™ Detergent Solution (Thermo Fisher Scientific, Inc.)]. .. The gels were subsequently stained with Coomassie Brilliant Blue R250 and destained until the wash remained clear and cleared zones associated with MMP activity were apparent.

Article Title: Dual Readout BRET/FRET Sensors for Measuring Intracellular Zinc
Article Snippet: Paragraph title: Mammalian Cell Culture and in Situ [Zn2+ ] Measurements in Cell Suspension ... The measurement was performed for 50 min, during which 50 μM N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN, Sigma-Aldrich) was loaded onto the cells after 15 min and 100 μM ZnCl2 and 5 μM 2-mercaptopyridine N-oxide (pyrithione, Acros Organics) after 35 min. During the additions of reagents, the measurement was stopped for ∼1 min. Sensor occupancies were calculated using , and corresponding Zn2+ levels were calculated using , in which R min and R max are the steady-state emission ratios after the addition of TPEN and Zn2+ 2 3

Generated:

Article Title: Development and in vitro assessment of enzymatically-responsive poly(ethylene glycol) hydrogels for the delivery of therapeutic peptides
Article Snippet: Hydrogels were swollen in buffer (50 mM Tricine (Acros Organics), 50 mM NaCl, 10 mM CaCl2 , 50 μM ZnCl2 (Alfa Aesar), and 0.05 wt% Brij35 (Alfa Aesar) in ddH2 O, pH 7.4) at 37 °C for 24 hours to achieve equilibrium swelling before characterization. .. Hydrogels were swollen in buffer (50 mM Tricine (Acros Organics), 50 mM NaCl, 10 mM CaCl2 , 50 μM ZnCl2 (Alfa Aesar), and 0.05 wt% Brij35 (Alfa Aesar) in ddH2 O, pH 7.4) at 37 °C for 24 hours to achieve equilibrium swelling before characterization.

other:

Article Title: Histidine phosphorylation relieves copper inhibition in the mammalian potassium channel KCa3.1
Article Snippet: TPEN (Sigma-Aldrich, St. Louis, MO, Cat # P4413), TTM (Sigma-Aldrich, Cat # 323446), CuCl2 (Sigma-Aldrich, Cat # C6641) and ZnCl2 (Acros, Geel, Belgium, Cat # 318170100) were purchased.

Article Title: Effect of Magnesium Incorporation on Solution-Processed Kesterite Solar Cells
Article Snippet: The CZTSSe precursor solution contained thiourea (99%+, Sigma-Aldrich), SnCl2 ·2H2 O (98%, Sigma-Aldrich), ZnCl2 (99.99%, Alfa Aesar) and CuCl2 (98%+, Alfa Aesar) dissolved in DMSO (99.9%, Alfa Aesar).

Article Title: Characterization of Biochemical Properties of Bacillus subtilis RecQ Helicase
Article Snippet: Puratronic CaCl2 , MgCl2 , MnCl2 , NiCl2 , and ZnCl2 were from Alfa Aesar (Ward Hill, MA).

Imaging:

Article Title: Snapshot Hyperspectral Light-Sheet Imaging of Signal Transduction in Live Pancreatic Islets
Article Snippet: Three glucose concentrations in KRBH were used for the imaging experiments: 1, 2, and 10 mM glucose. .. ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4.

Protein Concentration:

Article Title: Inhibitory effect of norcantharidin on melanoma tumor growth and vasculogenic mimicry by suppressing MMP-2 expression
Article Snippet: Paragraph title: Zymography assays and MMP-2 protein concentration determination ... The gel was washed twice in 2.5% (w/v) Triton X-100 solution and incubated overnight at 37°C in developing buffer [50 mmol/l Tris/HCl (pH 7.4), 10 mmol/l CaCl2 , 5 mmol/l ZnCl2 and 0.05% Brij™ 35 Surfact-Amps™ Detergent Solution (Thermo Fisher Scientific, Inc.)].

Sequencing:

Article Title: Complete Genome Sequence of an Ebola Virus Isolate Imported from Sierra Leone to Germany Determined by Circle Sequencing
Article Snippet: We used a novel circle sequencing approach , a technology that allows highly accurate genome sequencing of RNA virus genomes, with optimization as described below. .. The resulting rRNA-free sample was fragmented with ZnCl2 for 11 min at 94°C (Ambion).

Sonication:

Article Title: Snapshot Hyperspectral Light-Sheet Imaging of Signal Transduction in Live Pancreatic Islets
Article Snippet: Briefly, 1 mM dye in dimethyl sulfoxide was diluted in 1 ml Krebs Ringer Bicarbonate HEPES buffer (KRBH) to a final concentration of 4 μ M. To disperse the hydrophobic dye molecules into solution, the final buffer were sonicated for 10 min. Islets were incubated in this buffer at room temperature for 45 min with gentle rocking. .. ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4.

Binding Assay:

Article Title: Snapshot Hyperspectral Light-Sheet Imaging of Signal Transduction in Live Pancreatic Islets
Article Snippet: ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4. .. ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4.

Nucleic Acid Electrophoresis:

Article Title: Exploring mRNA 3?-UTR G-quadruplexes: evidence of roles in both alternative polyadenylation and mRNA shortening
Article Snippet: To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen). .. To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen).

Article Title: The folding of 5′-UTR human G-quadruplexes possessing a long central loop
Article Snippet: In order to produce 5′-end-labeled RNA molecules, purified transcripts (50 pmol) were dephosphorylated at 37°C for 30 min by adding 5 units of antarctic phosphatase (New England BioLabs) in a final volume of 10 μL containing 50 mM Bis-propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 , and 20 units RNase OUT (Invitrogen). .. In order to produce 5′-end-labeled RNA molecules, purified transcripts (50 pmol) were dephosphorylated at 37°C for 30 min by adding 5 units of antarctic phosphatase (New England BioLabs) in a final volume of 10 μL containing 50 mM Bis-propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 , and 20 units RNase OUT (Invitrogen).

Fluorescence:

Article Title: Snapshot Hyperspectral Light-Sheet Imaging of Signal Transduction in Live Pancreatic Islets
Article Snippet: ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4. .. ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4.

Isolation:

Article Title: Complete Genome Sequence of an Ebola Virus Isolate Imported from Sierra Leone to Germany Determined by Circle Sequencing
Article Snippet: The virus was isolated from a highly positive serum sample (day 9) and passaged two times each in Vero E6 and HuH7 cells for virus propagation (sequences from p4 stocks). .. The resulting rRNA-free sample was fragmented with ZnCl2 for 11 min at 94°C (Ambion).

Labeling:

Article Title: CRISPRscan: designing highly efficient sgRNAs for CRISPR/Cas9 targeting in vivo
Article Snippet: Paragraph title: sgRNA labeling and in-line probing ... To produce 5′-end-labeled sgRNAs, purified sgRNA transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BIoLabs) to 50 pmol of sgRNA in a final volume of 10 μL containing 50 mM Bis-Propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20U, Invitrogen).

Article Title: Snapshot Hyperspectral Light-Sheet Imaging of Signal Transduction in Live Pancreatic Islets
Article Snippet: ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4. .. ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4.

Article Title: Exploring mRNA 3?-UTR G-quadruplexes: evidence of roles in both alternative polyadenylation and mRNA shortening
Article Snippet: Paragraph title: RNA synthesis and labeling ... To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen).

Article Title: 5?-UTR G-quadruplex structures acting as translational repressors
Article Snippet: Paragraph title: RNA labeling ... In order to produce 5′-end-labeled PG4s, purified transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BioLabs,) to 50 pmol of RNA and incubating the reaction mixture for 30 min at 37°C in a final volume of 10 µl containing 50 mM Bis-Propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen).

Article Title: 3′ UTR G-quadruplexes regulate miRNA binding
Article Snippet: Paragraph title: RNA labeling ... Purified transcripts (50 pmol) were dephosphorylated for 1 h at 37°C in a solution containing 1 U of Antarctic phosphatase (New England Biolabs), 50 mM bis-propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 , and RNase OUT (20 U, Invitrogen) in a final volume of 10 µL.

Purification:

Article Title: CRISPRscan: designing highly efficient sgRNAs for CRISPR/Cas9 targeting in vivo
Article Snippet: DNA was then eluted, ethanol precipitated, dissolved in water and PCR amplified as mentioned above. .. To produce 5′-end-labeled sgRNAs, purified sgRNA transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BIoLabs) to 50 pmol of sgRNA in a final volume of 10 μL containing 50 mM Bis-Propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20U, Invitrogen). .. Dephosphorylated sgRNAs (5 pmol) were 5′-end-rediolabeled using 3 U of T4 polynucleotide kinase (New England Biolabs) for 1 h at 37°C in the presence of 3.2 pmol of [α-32 P]ATP (6000 Ci/mmol; Perkin Elmer).

Article Title: Complete Genome Sequence of an Ebola Virus Isolate Imported from Sierra Leone to Germany Determined by Circle Sequencing
Article Snippet: The resulting rRNA-free sample was fragmented with ZnCl2 for 11 min at 94°C (Ambion). .. The resulting rRNA-free sample was fragmented with ZnCl2 for 11 min at 94°C (Ambion).

Article Title: New scoring system to identify RNA G-quadruplex folding
Article Snippet: Melting temperature values (Tm ) were calculated using ‘fraction folded’ (θ) versus temperature plots ( ). .. Before radioactive 5′-end-labelling, 50 pmol of purified RNA transcripts were dephosphorylated using 1 U of antarctic phosphatase (New England BioLabs) in a 10-µl final volume reaction containing 50 mM Bis-Tris-propane, pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and 20 U RNase OUT (Invitrogen). .. Reactions were incubated at 37°C for 30 min and the enzyme was then inactivated by incubating at 65°C for 7 min. For the 5′-end-labelling reaction itself, dephosphorylated transcripts (10 pmol) were incubated at 37°C for 1 h in the presence of 3 U of T4 polynucleotide kinase (USB), 3.2 pmol of [γ-32 P] ATP (6000 Ci/mmol; New England Nuclear), 20 U of RNase OUT (Invitrogen) and in a buffer with final concentrations of 50 mM Tris–HCl, pH 7.6, 10 mM MgCl2, 10 mM β-mercaptoethanol, all in a final volume of 10 µl.

Article Title: Exploring mRNA 3?-UTR G-quadruplexes: evidence of roles in both alternative polyadenylation and mRNA shortening
Article Snippet: The PG4s were then ethanol precipitated, dried, dissolved in water and analyzed by spectrometry at 260 nm to determine their concentration. .. To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen). .. Dephosphorylated RNAs (5 pmol) were 5′-end radiolabeled using 3 U of T4 polynucleotide kinase (Promega) for 1 h at 37°C in the presence of 3.2 pmol of [γ-32 P]adenosine triphosphate (6000 Ci/mmol; New England Nuclear).

Article Title: The Importance of Peripheral Sequences in Determining the Metal Selectivity of an in Vitro-Selected Co2+-Dependent DNAzyme
Article Snippet: NaCl, CoCl2 , ZnCl2 and Pb(CH3 COO)2 were purchased from Alfa Aesar at puratronic grade (99.998% or greater purity, metals basis). .. HPLC-purified DNA oligonucleotides were purchased from Integrated DNA Technologies.

Article Title: 5?-UTR G-quadruplex structures acting as translational repressors
Article Snippet: CD melting curves were obtained by heating the samples from 25°C to 90°C at a controlled rate of 1°C min−1 and monitoring a 264-nm CD peak every 0.2 min. Melting temperature (T m ) values were calculated using ‘fraction folded’ (θ ) versus temperature plots ( ). .. In order to produce 5′-end-labeled PG4s, purified transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BioLabs,) to 50 pmol of RNA and incubating the reaction mixture for 30 min at 37°C in a final volume of 10 µl containing 50 mM Bis-Propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen). .. Dephosphorylated transcripts (5 pmol) were 5′-end-radiolabeled using 3 U of T4 polynucleotide kinase (Promega) for 1 h at 37°C in the presence of 3.2 pmol of [α-32 P]ATP (6000 Ci/mmol; New England Nuclear).

Article Title: 3′ UTR G-quadruplexes regulate miRNA binding
Article Snippet: The RNA was then quantified by absorbance at 260 nm. .. Purified transcripts (50 pmol) were dephosphorylated for 1 h at 37°C in a solution containing 1 U of Antarctic phosphatase (New England Biolabs), 50 mM bis-propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 , and RNase OUT (20 U, Invitrogen) in a final volume of 10 µL. .. Dephosphorylated transcripts (5 pmol) were 5′-end-radiolabeled using 3 U of T4 polynucleotide kinase (Promega) for 1 h at 37°C in the presence of 3.2 pmol of [α-32 P]ATP (6000 Ci/mmol; New England Nuclear) in a final volume of 10 µL.

Article Title: The folding of 5′-UTR human G-quadruplexes possessing a long central loop
Article Snippet: The concentrations were determined by spectrometry at 260 nm using a NanoVue system (GE Healthcare). .. In order to produce 5′-end-labeled RNA molecules, purified transcripts (50 pmol) were dephosphorylated at 37°C for 30 min by adding 5 units of antarctic phosphatase (New England BioLabs) in a final volume of 10 μL containing 50 mM Bis-propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 , and 20 units RNase OUT (Invitrogen). .. The dephosphorylated RNAs (10 pmol) were 5′-end-radiolabeled using 7.5 units of T4 polynucleotide kinase (Promega) for 1 h at 37°C in the presence of 3.2 pmol of [γ-32 P]ATP (6000 Ci/mmol; New England Nuclear).

Polyacrylamide Gel Electrophoresis:

Article Title: CRISPRscan: designing highly efficient sgRNAs for CRISPR/Cas9 targeting in vivo
Article Snippet: To produce 5′-end-labeled sgRNAs, purified sgRNA transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BIoLabs) to 50 pmol of sgRNA in a final volume of 10 μL containing 50 mM Bis-Propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20U, Invitrogen). .. To produce 5′-end-labeled sgRNAs, purified sgRNA transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BIoLabs) to 50 pmol of sgRNA in a final volume of 10 μL containing 50 mM Bis-Propane pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20U, Invitrogen).

Article Title: New scoring system to identify RNA G-quadruplex folding
Article Snippet: Before radioactive 5′-end-labelling, 50 pmol of purified RNA transcripts were dephosphorylated using 1 U of antarctic phosphatase (New England BioLabs) in a 10-µl final volume reaction containing 50 mM Bis-Tris-propane, pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and 20 U RNase OUT (Invitrogen). .. Before radioactive 5′-end-labelling, 50 pmol of purified RNA transcripts were dephosphorylated using 1 U of antarctic phosphatase (New England BioLabs) in a 10-µl final volume reaction containing 50 mM Bis-Tris-propane, pH 6.0, 1 mM MgCl2 , 0.1 mM ZnCl2 and 20 U RNase OUT (Invitrogen).

Article Title: Exploring mRNA 3?-UTR G-quadruplexes: evidence of roles in both alternative polyadenylation and mRNA shortening
Article Snippet: RNA products were fractionated by denaturing (8 M urea) 10% polyacrylamide gel electrophoresis (PAGE; 19:1 ratio of acrylamide to bisacrylamide) using 45 mM Tris–borate, pH 7.5, and 1 mM ethylenediaminetetraacetic acid (EDTA) solution as running buffer. .. To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen).

Article Title: 5?-UTR G-quadruplex structures acting as translational repressors
Article Snippet: In order to produce 5′-end-labeled PG4s, purified transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BioLabs,) to 50 pmol of RNA and incubating the reaction mixture for 30 min at 37°C in a final volume of 10 µl containing 50 mM Bis-Propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen). .. In order to produce 5′-end-labeled PG4s, purified transcripts were dephosphorylated by adding 1 U of antartic phosphatase (New England BioLabs,) to 50 pmol of RNA and incubating the reaction mixture for 30 min at 37°C in a final volume of 10 µl containing 50 mM Bis-Propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen).

Article Title: 3′ UTR G-quadruplexes regulate miRNA binding
Article Snippet: Purified transcripts (50 pmol) were dephosphorylated for 1 h at 37°C in a solution containing 1 U of Antarctic phosphatase (New England Biolabs), 50 mM bis-propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 , and RNase OUT (20 U, Invitrogen) in a final volume of 10 µL. .. Purified transcripts (50 pmol) were dephosphorylated for 1 h at 37°C in a solution containing 1 U of Antarctic phosphatase (New England Biolabs), 50 mM bis-propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 , and RNase OUT (20 U, Invitrogen) in a final volume of 10 µL.

Titration:

Article Title: Engineering glycoside hydrolase stability by the introduction of zinc binding
Article Snippet: DSF assays comparing the thermal stability of J30 wt and J30 CCH were performed on an Applied Biosciences StepOnePlus Real-Time PCR System in triplicate on a 30 µl scale containing 0.5 mg ml−1 enzyme, 150 mM NaCl, 25 mM HEPES pH 7.4, 10 µM ZnCl2 and SYPRO Orange Protein Gel Stain (Life Technologies, catalog No. S-6650) at 5×. .. DSF assays comparing the thermal stability of J30 wt and J30 CCH were performed on an Applied Biosciences StepOnePlus Real-Time PCR System in triplicate on a 30 µl scale containing 0.5 mg ml−1 enzyme, 150 mM NaCl, 25 mM HEPES pH 7.4, 10 µM ZnCl2 and SYPRO Orange Protein Gel Stain (Life Technologies, catalog No. S-6650) at 5×.

SDS Page:

Article Title: Inhibitory effect of norcantharidin on melanoma tumor growth and vasculogenic mimicry by suppressing MMP-2 expression
Article Snippet: Gelatin zymography was used to examine the levels of MMP-2 activity in A375 cells that were either untreated (control group) or treated with 28 µg/ml NTCD for 48 h. The culture media were collected and subjected to 10% SDS-PAGE using 0.01% w/v gelatin. .. The gel was washed twice in 2.5% (w/v) Triton X-100 solution and incubated overnight at 37°C in developing buffer [50 mmol/l Tris/HCl (pH 7.4), 10 mmol/l CaCl2 , 5 mmol/l ZnCl2 and 0.05% Brij™ 35 Surfact-Amps™ Detergent Solution (Thermo Fisher Scientific, Inc.)].

Plasmid Preparation:

Article Title: Interaction of catechol and non-catechol substrates with externally or internally facing dopamine transporters
Article Snippet: The pCIN4 vector and the pCIN4-synthetic human DAT construct (pCIN4-DAT) were generous gifts from Dr. Jonathan A. Javitch (Columbia University, New York, NY, USA). .. Digitonin and ZnCl2 were from Acros Organics Co. (Geel, Belgium).

Sample Prep:

Article Title: Snapshot Hyperspectral Light-Sheet Imaging of Signal Transduction in Live Pancreatic Islets
Article Snippet: Unlike many light-sheet imaging approaches, the iSPIM does not require complex sample preparation (i.e., agar embedding), but instead accommodates any specimen mounted on a standard coverslip. .. ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4.

In Situ:

Article Title: Dual Readout BRET/FRET Sensors for Measuring Intracellular Zinc
Article Snippet: Paragraph title: Mammalian Cell Culture and in Situ [Zn2+ ] Measurements in Cell Suspension ... The measurement was performed for 50 min, during which 50 μM N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN, Sigma-Aldrich) was loaded onto the cells after 15 min and 100 μM ZnCl2 and 5 μM 2-mercaptopyridine N-oxide (pyrithione, Acros Organics) after 35 min. During the additions of reagents, the measurement was stopped for ∼1 min. Sensor occupancies were calculated using , and corresponding Zn2+ levels were calculated using , in which R min and R max are the steady-state emission ratios after the addition of TPEN and Zn2+ 2 3

Ethanol Precipitation:

Article Title: Exploring mRNA 3?-UTR G-quadruplexes: evidence of roles in both alternative polyadenylation and mRNA shortening
Article Snippet: The RNA was then purified by phenol:chloroform extraction followed by ethanol precipitation. .. To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen).

Direct Antiglobulin Test:

Article Title: Interaction of catechol and non-catechol substrates with externally or internally facing dopamine transporters
Article Snippet: The pCIN4 vector and the pCIN4-synthetic human DAT construct (pCIN4-DAT) were generous gifts from Dr. Jonathan A. Javitch (Columbia University, New York, NY, USA). .. Digitonin and ZnCl2 were from Acros Organics Co. (Geel, Belgium).

Concentration Assay:

Article Title: Zn-based eutectic mixture as anolyte for hybrid redox flow batteries
Article Snippet: To prepare Zn eutectic solvent, acetamide (99%, Alfa Aesar) and ZnCl2 (anhydrous, 99.99%, Alfa Aesar) with different molar ratio were mixed under constant stirring at 60 °C to accelerate the formation of Zn eutectic solvent until a transparent liquid formed. .. To prepare Zn eutectic solvent, acetamide (99%, Alfa Aesar) and ZnCl2 (anhydrous, 99.99%, Alfa Aesar) with different molar ratio were mixed under constant stirring at 60 °C to accelerate the formation of Zn eutectic solvent until a transparent liquid formed.

Article Title: Snapshot Hyperspectral Light-Sheet Imaging of Signal Transduction in Live Pancreatic Islets
Article Snippet: Three glucose concentrations in KRBH were used for the imaging experiments: 1, 2, and 10 mM glucose. .. ZnCl2 (Acros Organics, Thermo Scientific) was diluted in KRBH, to a final concentration of 30 μ M. The pH of all solutions used was stabilized at 7.4. .. Standard image processing was performed on the images after the linear unmixing.

Article Title: Exploring mRNA 3?-UTR G-quadruplexes: evidence of roles in both alternative polyadenylation and mRNA shortening
Article Snippet: The PG4s were then ethanol precipitated, dried, dissolved in water and analyzed by spectrometry at 260 nm to determine their concentration. .. To produce 5′-end-labeled RNA molecules, 50 pmol of purified transcripts were dephosphorylated at 37°C during 1 h in the presence of 1 U of Antarctic phosphatase (New England BioLabs) in a final volume of 10 µl containing 50 mM Bis–propane (pH 6.0), 1 mM MgCl2 , 0.1 mM ZnCl2 and RNase OUT (20 U, Invitrogen).

Live Cell Imaging:

Article Title: Dual Readout BRET/FRET Sensors for Measuring Intracellular Zinc
Article Snippet: Two days after transfection, cells were released from the wells using trypsin (Thermo Fisher Scientific) and resuspended in 1 mL of preheated (37 °C) live-cell imaging buffer (20 mM HEPES (pH 7.4), 140 mM NaCl, 2.5 mM KCl, 1.8 mM CaCl2 , and 1.0 mM MgCl2 ). .. The measurement was performed for 50 min, during which 50 μM N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN, Sigma-Aldrich) was loaded onto the cells after 15 min and 100 μM ZnCl2 and 5 μM 2-mercaptopyridine N-oxide (pyrithione, Acros Organics) after 35 min. During the additions of reagents, the measurement was stopped for ∼1 min. Sensor occupancies were calculated using , and corresponding Zn2+ levels were calculated using , in which R min and R max are the steady-state emission ratios after the addition of TPEN and Zn2+ 2 3

Staining:

Article Title: Engineering glycoside hydrolase stability by the introduction of zinc binding
Article Snippet: The figures showing structural models were created using PyMOL (v.1.7.2.1; Schrödinger). .. DSF assays comparing the thermal stability of J30 wt and J30 CCH were performed on an Applied Biosciences StepOnePlus Real-Time PCR System in triplicate on a 30 µl scale containing 0.5 mg ml−1 enzyme, 150 mM NaCl, 25 mM HEPES pH 7.4, 10 µM ZnCl2 and SYPRO Orange Protein Gel Stain (Life Technologies, catalog No. S-6650) at 5×. .. Fluorescence signals were recorded from 25 to 99°C.

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    Thermo Fisher zncl2
    Zncl2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    zncl2 - by Bioz Stars, 2019-10
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