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Millipore m cacl2
M Cacl2, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 96 stars, based on 7 article reviews
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m cacl2 - by Bioz Stars, 2020-04
96/100 stars

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Related Articles

Clone Assay:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Human CaM cDNA was subcloned into the pET-15b expression vector (Novagen, San Diego, CA) using standard molecular biology cloning techniques. .. Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA.

Centrifugation:

Article Title: Primary Role for Endoplasmic Reticulum-bound Ribosomes in Cellular Translation Identified by Ribosome Profiling *
Article Snippet: Cell fractions were adjusted to 100 m m KOAc, 25 m m K-HEPES, pH 7.2, 15 m m Mg(OAc)2 , 3 m m CaCl2 , 180 μ m cycloheximide and treated with 20 μg/ml micrococcal nuclease (Sigma-Aldrich) for 1 h at 37 °C ( ). .. Nuclease activity was inactivated by the addition of 6 m m EGTA, and ribosomes were recovered by centrifugation (60 min, 90,000 RPM in the Beckman TLA 100.3 rotor) over a 500 m m sucrose cushion.

Article Title: The Baseplate of Lactobacillus delbrueckii Bacteriophage Ld17 Harbors a Glycerophosphodiesterase *
Article Snippet: The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min. .. The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min.

Article Title: Nifetepimine, a Dihydropyrimidone, Ensures CD4+ T Cell Survival in a Tumor Microenvironment by Maneuvering Sarco(endo)plasmic Reticulum Ca2+ ATPase (SERCA) *
Article Snippet: After centrifugation, 0.5 to 1 × 106 cells were suspended in 3 ml of Hanks' buffered saline solution in a quartz cuvette and inserted into a Hitachi spectrofluorimeter equipped with a stirring apparatus and a thermostated (37 °C) cuvette holder and connected to a PC computer. .. Maximum Fura-2 fluorescence (Fmax) was obtained by adding 1 μ m ionomycin (Sigma) to the cell suspension in the presence of 10 m m CaCl2 , and minimum fluorescence (Fmin) was determined without added calcium in the presence of 5 m m EGTA (Sigma).

Article Title: Redox Regulation of Transglutaminase 2 Activity *
Article Snippet: .. The components were as follows: 1) a concentrated buffer consisting of 1 m MOPS, 5 m m EDTA, and 50 m m α-ketoglutarate (pH 7.2); 2) 100 m m CaCl2 ; 3) 300 m m benzyloxycarbonyl-Gln-Gly (Sigma) prepared as a solution of the sodium salt by titration with sodium hydroxide and diluted to 10× stocks of the desired reaction concentrations; 4) 50 m m NADH; and 5) lyophilized ammonium-free glutamate dehydrogenase (Biozyme Laboratories) dissolved to 0.5 units/μl in 200 m m MOPS, 1 m m EDTA, and 10 m m α-ketoglutarate (pH 7.2) and clarified by centrifugation. ..

Neurite Outgrowth Assay:

Article Title: Neuronal Matrix Metalloproteinase-2 Degrades and Inactivates a Neurite-Inhibiting Chondroitin Sulfate Proteoglycan
Article Snippet: Cryoculture is a neurite outgrowth assay in which neurons are cultured directly on unfixed nerve sections ( ). .. Before cryoculture, mounted sections were treated for 4 hr at 37°C with (1) activated MMP-2 (10 μg/ml) in 25 m m Tris-HCl, pH 7.8, containing 2.5 m m CaCl2 ; (2) chondroitinase ABC (0.1 U/ml) (Sigma) in 50 m m Tris-HCl, pH 8.0, containing 50 m m NaCl; or (3) buffer control.

Construct:

Article Title: The Mechanoenzymatic Core of Dynamin-related Protein 1 Comprises the Minimal Machinery Required for Membrane Constriction *
Article Snippet: The Drp1 ΔVD and K38A construct were made using the QuikChange mutagenesis kit (Agilent Technologies). .. The overnight culture was harvested, and cell pellets were resuspended in a cell lysis/binding buffer (500 m m arginine, pH 8, 300 m m NaCl, 10 m m β-mercaptoethanol (BME), 5.0 m m magnesium chloride, 1.0 m m imidazole, and 2 m m CaCl2 ) with 100 ng/μl lysosome (Sigma), 4 units/ml DNase (Sigma), and complete EDTA-free protease inhibitor mixture tablets (Roche).

Electrophoresis:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA. .. CaM purity was greater than 95% as judged by SDS gel electrophoresis and Coomassie Brilliant Blue staining.

Incubation:

Article Title: Enhanced Interferon Signaling Pathway in Oral Cancer Revealed by Quantitative Proteome Analysis of Microdissected Specimens Using 16O/18O Labeling and Integrated Two-dimensional LC-ESI-MALDI Tandem MS *
Article Snippet: After drying the peptides under a vacuum, the samples were added to 16 O or 18 O labeling solutions containing 1 μg of trypsin, 20 m m CaCl2 , and 50 m m Tris-HCl, pH 8.0, in H2 16 O or H2 18 O (Sigma-Aldrich). .. Trypsin was inactivated by incubation in a boiling water bath for 15 min and acidification with formic acid to a final concentration of 3% (similar to that described in a previous report ( )).

Article Title: Primary Role for Endoplasmic Reticulum-bound Ribosomes in Cellular Translation Identified by Ribosome Profiling *
Article Snippet: Cell fractions were adjusted to 100 m m KOAc, 25 m m K-HEPES, pH 7.2, 15 m m Mg(OAc)2 , 3 m m CaCl2 , 180 μ m cycloheximide and treated with 20 μg/ml micrococcal nuclease (Sigma-Aldrich) for 1 h at 37 °C ( ). .. The purified ribosome pellet was resuspended in 50 m m sodium chloride, 50 m m Tris, pH 7.5, 5 m m EDTA, 0.5% SDS, 200 μg/ml proteinase K and incubated for 30 min at 37 °C ( ).

Article Title: The Stoichiometry and Biophysical Properties of the Kv4 Potassium Channel Complex with K+ Channel-interacting Protein (KChIP) Subunits Are Variable, Depending on the Relative Expression Level *
Article Snippet: .. The injected oocytes were incubated for 1–3 days at 17 °C in frog Ringer solution containing 88 m m NaCl, 1 m m KCl, 2.4 m m NaHCO3 , 0.3 m m Ca(NO3 )2 , 0.41 m m CaCl2 , and 0.82 m m MgSO4 , pH 7.6, with 0.1% penicillin-streptomycin solution (Sigma-Aldrich). ..

Article Title: The Baseplate of Lactobacillus delbrueckii Bacteriophage Ld17 Harbors a Glycerophosphodiesterase *
Article Snippet: .. The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min. .. The bacterial suspension was subsequently placed on ice and disrupted with glass beads in a Mini Bead Beater (Biospec Products, Bartlesville, OK) for 90 s followed by 5 min on ice for three cycles.

Article Title: Nifetepimine, a Dihydropyrimidone, Ensures CD4+ T Cell Survival in a Tumor Microenvironment by Maneuvering Sarco(endo)plasmic Reticulum Ca2+ ATPase (SERCA) *
Article Snippet: Cells were centrifuged and resuspended at a density of 106 cells/ml in PBS supplemented with 1 mg/ml bovine serum albumin and incubated in the dark with Fura-2AM (final concentration, 5 μ m ) (Sigma) for 30 min at room temperature under slow agitation. .. Maximum Fura-2 fluorescence (Fmax) was obtained by adding 1 μ m ionomycin (Sigma) to the cell suspension in the presence of 10 m m CaCl2 , and minimum fluorescence (Fmin) was determined without added calcium in the presence of 5 m m EGTA (Sigma).

Activity Assay:

Article Title: Primary Role for Endoplasmic Reticulum-bound Ribosomes in Cellular Translation Identified by Ribosome Profiling *
Article Snippet: Cell fractions were adjusted to 100 m m KOAc, 25 m m K-HEPES, pH 7.2, 15 m m Mg(OAc)2 , 3 m m CaCl2 , 180 μ m cycloheximide and treated with 20 μg/ml micrococcal nuclease (Sigma-Aldrich) for 1 h at 37 °C ( ). .. Nuclease activity was inactivated by the addition of 6 m m EGTA, and ribosomes were recovered by centrifugation (60 min, 90,000 RPM in the Beckman TLA 100.3 rotor) over a 500 m m sucrose cushion.

Solubility:

Article Title: The Baseplate of Lactobacillus delbrueckii Bacteriophage Ld17 Harbors a Glycerophosphodiesterase *
Article Snippet: Protein expression using the nisin-inducible expression system of pTX8048 provides an N-terminal protein fusion to thioredoxin to improve protein solubility and to a His6 tag for protein purification purposes ( ). .. The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min.

Expressing:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Paragraph title: CaM Expression and Purification ... Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA.

Article Title: The Mechanoenzymatic Core of Dynamin-related Protein 1 Comprises the Minimal Machinery Required for Membrane Constriction *
Article Snippet: Paragraph title: Drp1 Expression and Purification ... The overnight culture was harvested, and cell pellets were resuspended in a cell lysis/binding buffer (500 m m arginine, pH 8, 300 m m NaCl, 10 m m β-mercaptoethanol (BME), 5.0 m m magnesium chloride, 1.0 m m imidazole, and 2 m m CaCl2 ) with 100 ng/μl lysosome (Sigma), 4 units/ml DNase (Sigma), and complete EDTA-free protease inhibitor mixture tablets (Roche).

Article Title: The Baseplate of Lactobacillus delbrueckii Bacteriophage Ld17 Harbors a Glycerophosphodiesterase *
Article Snippet: Paragraph title: Recombinant Protein Expression and Purification ... The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min.

Acrylamide Gel Assay:

Article Title: Primary Role for Endoplasmic Reticulum-bound Ribosomes in Cellular Translation Identified by Ribosome Profiling *
Article Snippet: Cell fractions were adjusted to 100 m m KOAc, 25 m m K-HEPES, pH 7.2, 15 m m Mg(OAc)2 , 3 m m CaCl2 , 180 μ m cycloheximide and treated with 20 μg/ml micrococcal nuclease (Sigma-Aldrich) for 1 h at 37 °C ( ). .. RNA was purified by phenol/chloroform extraction and separated on a 15% acrylamide gel containing 1× Tris-borate-EDTA and 7 m urea.

Transformation Assay:

Article Title: The Mechanoenzymatic Core of Dynamin-related Protein 1 Comprises the Minimal Machinery Required for Membrane Constriction *
Article Snippet: Briefly, CBP-Drp1 was transformed into Escherichia coli BL21 DE3 cells. .. The overnight culture was harvested, and cell pellets were resuspended in a cell lysis/binding buffer (500 m m arginine, pH 8, 300 m m NaCl, 10 m m β-mercaptoethanol (BME), 5.0 m m magnesium chloride, 1.0 m m imidazole, and 2 m m CaCl2 ) with 100 ng/μl lysosome (Sigma), 4 units/ml DNase (Sigma), and complete EDTA-free protease inhibitor mixture tablets (Roche).

Chromatography:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA. .. The identity of the purified CaM was confirmed by N-terminal sequencing and by liquid chromatography-mass spectrometry analysis of its tryptic digest fragments.

Protease Inhibitor:

Article Title: The Mechanoenzymatic Core of Dynamin-related Protein 1 Comprises the Minimal Machinery Required for Membrane Constriction *
Article Snippet: .. The overnight culture was harvested, and cell pellets were resuspended in a cell lysis/binding buffer (500 m m arginine, pH 8, 300 m m NaCl, 10 m m β-mercaptoethanol (BME), 5.0 m m magnesium chloride, 1.0 m m imidazole, and 2 m m CaCl2 ) with 100 ng/μl lysosome (Sigma), 4 units/ml DNase (Sigma), and complete EDTA-free protease inhibitor mixture tablets (Roche). .. The resuspended cells were then passed through a micro fluidizer (M-110 Y; Microfluidics, Newton, MA), and the lysate was centrifuged in a Beckman Coulter Optima L-look Ultracentrifuge (50.2 Ti rotor) at 40,000 rpm (184,048 × g ) for 30 min at 4 °C.

Article Title: Neuronal Matrix Metalloproteinase-2 Degrades and Inactivates a Neurite-Inhibiting Chondroitin Sulfate Proteoglycan
Article Snippet: The protease inhibitor phenanthroline initially was dissolved at 0.5 m in 50% ethanol and used at a final concentration of 1 m m . .. Before cryoculture, mounted sections were treated for 4 hr at 37°C with (1) activated MMP-2 (10 μg/ml) in 25 m m Tris-HCl, pH 7.8, containing 2.5 m m CaCl2 ; (2) chondroitinase ABC (0.1 U/ml) (Sigma) in 50 m m Tris-HCl, pH 8.0, containing 50 m m NaCl; or (3) buffer control.

Transferring:

Article Title: Postsynaptic Density 95 controls AMPA Receptor Incorporation during Long-Term Potentiation and Experience-Driven Synaptic Plasticity
Article Snippet: The peptide pep2m/G10 (KRMKVAKNAQ, Research Genetics, Huntsville, AL) was dissolved in internal solution (concentration, 2 m m ) and included in the patch pipette. .. AMPAR-mediated miniature currents (minis) were recorded at –60 mV in ACSF containing 4 m m CaCl2 , 2 m m MgCl2 , and 1 μ m TTX (Calbiochem).

Cell Culture:

Article Title: Neuronal Matrix Metalloproteinase-2 Degrades and Inactivates a Neurite-Inhibiting Chondroitin Sulfate Proteoglycan
Article Snippet: Cryoculture is a neurite outgrowth assay in which neurons are cultured directly on unfixed nerve sections ( ). .. Before cryoculture, mounted sections were treated for 4 hr at 37°C with (1) activated MMP-2 (10 μg/ml) in 25 m m Tris-HCl, pH 7.8, containing 2.5 m m CaCl2 ; (2) chondroitinase ABC (0.1 U/ml) (Sigma) in 50 m m Tris-HCl, pH 8.0, containing 50 m m NaCl; or (3) buffer control.

Generated:

Article Title: The Interaction of Integrin αIIbβ3 with Fibrin Occurs through Multiple Binding Sites in the αIIb β-Propeller Domain *
Article Snippet: Briefly, the reaction mixture (total volume, 1.0 ml) consisted of 3 × 108 platelets in isotonic HEPES buffer (20 m m HEPES, pH 7.3, 137 m m NaCl, 2.7 m m KCl, 1 m m MgCl2 , 3.3 m m NaH2 PO4 containing 35 mg/ml BSA and 1 mg/ml glucose), 0.25 mg/ml fibrinogen, and 1 m m CaCl2 in glass tubes coated with Sigmacote (Sigma). .. Clot retraction mediated by wild-type HEK293 cells and generated mutant cell lines was performed as described ( ).

Sequencing:

Article Title: Primary Role for Endoplasmic Reticulum-bound Ribosomes in Cellular Translation Identified by Ribosome Profiling *
Article Snippet: Paragraph title: Ribosome Footprint Preparation, Sequencing, and Mapping ... Cell fractions were adjusted to 100 m m KOAc, 25 m m K-HEPES, pH 7.2, 15 m m Mg(OAc)2 , 3 m m CaCl2 , 180 μ m cycloheximide and treated with 20 μg/ml micrococcal nuclease (Sigma-Aldrich) for 1 h at 37 °C ( ).

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA. .. The identity of the purified CaM was confirmed by N-terminal sequencing and by liquid chromatography-mass spectrometry analysis of its tryptic digest fragments.

Injection:

Article Title: The Stoichiometry and Biophysical Properties of the Kv4 Potassium Channel Complex with K+ Channel-interacting Protein (KChIP) Subunits Are Variable, Depending on the Relative Expression Level *
Article Snippet: .. The injected oocytes were incubated for 1–3 days at 17 °C in frog Ringer solution containing 88 m m NaCl, 1 m m KCl, 2.4 m m NaHCO3 , 0.3 m m Ca(NO3 )2 , 0.41 m m CaCl2 , and 0.82 m m MgSO4 , pH 7.6, with 0.1% penicillin-streptomycin solution (Sigma-Aldrich). ..

Recombinant:

Article Title: The Interaction of Integrin αIIbβ3 with Fibrin Occurs through Multiple Binding Sites in the αIIb β-Propeller Domain *
Article Snippet: Briefly, the reaction mixture (total volume, 1.0 ml) consisted of 3 × 108 platelets in isotonic HEPES buffer (20 m m HEPES, pH 7.3, 137 m m NaCl, 2.7 m m KCl, 1 m m MgCl2 , 3.3 m m NaH2 PO4 containing 35 mg/ml BSA and 1 mg/ml glucose), 0.25 mg/ml fibrinogen, and 1 m m CaCl2 in glass tubes coated with Sigmacote (Sigma). .. For clot retraction assays with recombinant Fg γ407, the volume of the reaction mixture was reduced to 0.25 ml.

Article Title: The Baseplate of Lactobacillus delbrueckii Bacteriophage Ld17 Harbors a Glycerophosphodiesterase *
Article Snippet: Paragraph title: Recombinant Protein Expression and Purification ... The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min.

Fluorescence:

Article Title: Nifetepimine, a Dihydropyrimidone, Ensures CD4+ T Cell Survival in a Tumor Microenvironment by Maneuvering Sarco(endo)plasmic Reticulum Ca2+ ATPase (SERCA) *
Article Snippet: .. Maximum Fura-2 fluorescence (Fmax) was obtained by adding 1 μ m ionomycin (Sigma) to the cell suspension in the presence of 10 m m CaCl2 , and minimum fluorescence (Fmin) was determined without added calcium in the presence of 5 m m EGTA (Sigma). .. The cytosolic Ca2+ was calculated from the Fura-2AM fluorescence intensity as [Ca2+ ]cyt = Kd ( F − F min)/( F max − F ), where Kd = 224 n m for Fura-2, and R is the ratio of fluorescence values ( F ) ( R = F340/F380).

Mutagenesis:

Article Title: The Interaction of Integrin αIIbβ3 with Fibrin Occurs through Multiple Binding Sites in the αIIb β-Propeller Domain *
Article Snippet: Briefly, the reaction mixture (total volume, 1.0 ml) consisted of 3 × 108 platelets in isotonic HEPES buffer (20 m m HEPES, pH 7.3, 137 m m NaCl, 2.7 m m KCl, 1 m m MgCl2 , 3.3 m m NaH2 PO4 containing 35 mg/ml BSA and 1 mg/ml glucose), 0.25 mg/ml fibrinogen, and 1 m m CaCl2 in glass tubes coated with Sigmacote (Sigma). .. Clot retraction mediated by wild-type HEK293 cells and generated mutant cell lines was performed as described ( ).

Article Title: The Mechanoenzymatic Core of Dynamin-related Protein 1 Comprises the Minimal Machinery Required for Membrane Constriction *
Article Snippet: The Drp1 ΔVD and K38A construct were made using the QuikChange mutagenesis kit (Agilent Technologies). .. The overnight culture was harvested, and cell pellets were resuspended in a cell lysis/binding buffer (500 m m arginine, pH 8, 300 m m NaCl, 10 m m β-mercaptoethanol (BME), 5.0 m m magnesium chloride, 1.0 m m imidazole, and 2 m m CaCl2 ) with 100 ng/μl lysosome (Sigma), 4 units/ml DNase (Sigma), and complete EDTA-free protease inhibitor mixture tablets (Roche).

Isolation:

Article Title: The Interaction of Integrin αIIbβ3 with Fibrin Occurs through Multiple Binding Sites in the αIIb β-Propeller Domain *
Article Snippet: Clot retraction assays using isolated platelets were performed as described previously ( ). .. Briefly, the reaction mixture (total volume, 1.0 ml) consisted of 3 × 108 platelets in isotonic HEPES buffer (20 m m HEPES, pH 7.3, 137 m m NaCl, 2.7 m m KCl, 1 m m MgCl2 , 3.3 m m NaH2 PO4 containing 35 mg/ml BSA and 1 mg/ml glucose), 0.25 mg/ml fibrinogen, and 1 m m CaCl2 in glass tubes coated with Sigmacote (Sigma).

Article Title: Postsynaptic Density 95 controls AMPA Receptor Incorporation during Long-Term Potentiation and Experience-Driven Synaptic Plasticity
Article Snippet: AMPAR-mediated miniature currents (minis) were recorded at –60 mV in ACSF containing 4 m m CaCl2 , 2 m m MgCl2 , and 1 μ m TTX (Calbiochem). .. NMDAR-mediated minis were isolated at –60 mV in ACSF containing 2 m m CaCl2 , no Mg2+ , 5 μ m NBQX (Sigma), and 1 μ m TTX.

Size-exclusion Chromatography:

Article Title: Postsynaptic Density 95 controls AMPA Receptor Incorporation during Long-Term Potentiation and Experience-Driven Synaptic Plasticity
Article Snippet: LTP was induced by pairing 3 Hz stimulation with depolarization of the postsynaptic neuron to 0 mV for 90 sec; recordings were maintained for at least 35 min after pairing. .. AMPAR-mediated miniature currents (minis) were recorded at –60 mV in ACSF containing 4 m m CaCl2 , 2 m m MgCl2 , and 1 μ m TTX (Calbiochem).

Labeling:

Article Title: Enhanced Interferon Signaling Pathway in Oral Cancer Revealed by Quantitative Proteome Analysis of Microdissected Specimens Using 16O/18O Labeling and Integrated Two-dimensional LC-ESI-MALDI Tandem MS *
Article Snippet: .. After drying the peptides under a vacuum, the samples were added to 16 O or 18 O labeling solutions containing 1 μg of trypsin, 20 m m CaCl2 , and 50 m m Tris-HCl, pH 8.0, in H2 16 O or H2 18 O (Sigma-Aldrich). .. Trypsin-catalyzed 16 O or 18 O labeling was performed overnight at 37 °C.

Purification:

Article Title: Complement activation by apoptotic endothelial cells following hypoxia/reoxygenation
Article Snippet: The following buffers were used: GVB (0·1% gelatin, 5 m m Veronal-buffered saline, pH 7·4), GVB+ + (GVB, 0·5 m m MgCl2 , 0·15 m m CaCl2 ), DGVB+ + (GVB diluted 1 : 1 with 5% dextrose, 0·5 m m MgCl2 , 0·15 m m CaCl2 ), and HBSS (Hanks' balanced salt solution, Sigma, St Louis, MO). .. DHS was reconstituted for the classical and alternative pathways by addition of 100 µg/ml purified C1q (Sigma) or 5 µg/ml purified factor D. Depletion and reconstitution of the classical and alternative pathways was confirmed by haemolytic assays.

Article Title: Primary Role for Endoplasmic Reticulum-bound Ribosomes in Cellular Translation Identified by Ribosome Profiling *
Article Snippet: Cell fractions were adjusted to 100 m m KOAc, 25 m m K-HEPES, pH 7.2, 15 m m Mg(OAc)2 , 3 m m CaCl2 , 180 μ m cycloheximide and treated with 20 μg/ml micrococcal nuclease (Sigma-Aldrich) for 1 h at 37 °C ( ). .. The purified ribosome pellet was resuspended in 50 m m sodium chloride, 50 m m Tris, pH 7.5, 5 m m EDTA, 0.5% SDS, 200 μg/ml proteinase K and incubated for 30 min at 37 °C ( ).

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Paragraph title: CaM Expression and Purification ... Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA.

Article Title: The Mechanoenzymatic Core of Dynamin-related Protein 1 Comprises the Minimal Machinery Required for Membrane Constriction *
Article Snippet: Paragraph title: Drp1 Expression and Purification ... The overnight culture was harvested, and cell pellets were resuspended in a cell lysis/binding buffer (500 m m arginine, pH 8, 300 m m NaCl, 10 m m β-mercaptoethanol (BME), 5.0 m m magnesium chloride, 1.0 m m imidazole, and 2 m m CaCl2 ) with 100 ng/μl lysosome (Sigma), 4 units/ml DNase (Sigma), and complete EDTA-free protease inhibitor mixture tablets (Roche).

Article Title: The Baseplate of Lactobacillus delbrueckii Bacteriophage Ld17 Harbors a Glycerophosphodiesterase *
Article Snippet: Paragraph title: Recombinant Protein Expression and Purification ... The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min.

Protein Purification:

Article Title: The Baseplate of Lactobacillus delbrueckii Bacteriophage Ld17 Harbors a Glycerophosphodiesterase *
Article Snippet: Protein expression using the nisin-inducible expression system of pTX8048 provides an N-terminal protein fusion to thioredoxin to improve protein solubility and to a His6 tag for protein purification purposes ( ). .. The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min.

Staining:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA. .. CaM purity was greater than 95% as judged by SDS gel electrophoresis and Coomassie Brilliant Blue staining.

Titration:

Article Title: Redox Regulation of Transglutaminase 2 Activity *
Article Snippet: .. The components were as follows: 1) a concentrated buffer consisting of 1 m MOPS, 5 m m EDTA, and 50 m m α-ketoglutarate (pH 7.2); 2) 100 m m CaCl2 ; 3) 300 m m benzyloxycarbonyl-Gln-Gly (Sigma) prepared as a solution of the sodium salt by titration with sodium hydroxide and diluted to 10× stocks of the desired reaction concentrations; 4) 50 m m NADH; and 5) lyophilized ammonium-free glutamate dehydrogenase (Biozyme Laboratories) dissolved to 0.5 units/μl in 200 m m MOPS, 1 m m EDTA, and 10 m m α-ketoglutarate (pH 7.2) and clarified by centrifugation. ..

Plasmid Preparation:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Human CaM cDNA was subcloned into the pET-15b expression vector (Novagen, San Diego, CA) using standard molecular biology cloning techniques. .. Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA.

Software:

Article Title: Postsynaptic Density 95 controls AMPA Receptor Incorporation during Long-Term Potentiation and Experience-Driven Synaptic Plasticity
Article Snippet: AMPAR-mediated miniature currents (minis) were recorded at –60 mV in ACSF containing 4 m m CaCl2 , 2 m m MgCl2 , and 1 μ m TTX (Calbiochem). .. Minis were analyzed using MiniAnalysis software (Synaptosoft, Decatur, GA).

SDS-Gel:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA. .. CaM purity was greater than 95% as judged by SDS gel electrophoresis and Coomassie Brilliant Blue staining.

Positron Emission Tomography:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: Human CaM cDNA was subcloned into the pET-15b expression vector (Novagen, San Diego, CA) using standard molecular biology cloning techniques. .. Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA.

Concentration Assay:

Article Title: Enhanced Interferon Signaling Pathway in Oral Cancer Revealed by Quantitative Proteome Analysis of Microdissected Specimens Using 16O/18O Labeling and Integrated Two-dimensional LC-ESI-MALDI Tandem MS *
Article Snippet: After drying the peptides under a vacuum, the samples were added to 16 O or 18 O labeling solutions containing 1 μg of trypsin, 20 m m CaCl2 , and 50 m m Tris-HCl, pH 8.0, in H2 16 O or H2 18 O (Sigma-Aldrich). .. Trypsin was inactivated by incubation in a boiling water bath for 15 min and acidification with formic acid to a final concentration of 3% (similar to that described in a previous report ( )).

Article Title: Nifetepimine, a Dihydropyrimidone, Ensures CD4+ T Cell Survival in a Tumor Microenvironment by Maneuvering Sarco(endo)plasmic Reticulum Ca2+ ATPase (SERCA) *
Article Snippet: Paragraph title: Determination of the Cytosolic Ca2+ Concentration ... Maximum Fura-2 fluorescence (Fmax) was obtained by adding 1 μ m ionomycin (Sigma) to the cell suspension in the presence of 10 m m CaCl2 , and minimum fluorescence (Fmin) was determined without added calcium in the presence of 5 m m EGTA (Sigma).

Article Title: Neuronal Matrix Metalloproteinase-2 Degrades and Inactivates a Neurite-Inhibiting Chondroitin Sulfate Proteoglycan
Article Snippet: The protease inhibitor phenanthroline initially was dissolved at 0.5 m in 50% ethanol and used at a final concentration of 1 m m . .. Before cryoculture, mounted sections were treated for 4 hr at 37°C with (1) activated MMP-2 (10 μg/ml) in 25 m m Tris-HCl, pH 7.8, containing 2.5 m m CaCl2 ; (2) chondroitinase ABC (0.1 U/ml) (Sigma) in 50 m m Tris-HCl, pH 8.0, containing 50 m m NaCl; or (3) buffer control.

Article Title: Postsynaptic Density 95 controls AMPA Receptor Incorporation during Long-Term Potentiation and Experience-Driven Synaptic Plasticity
Article Snippet: The peptide pep2m/G10 (KRMKVAKNAQ, Research Genetics, Huntsville, AL) was dissolved in internal solution (concentration, 2 m m ) and included in the patch pipette. .. AMPAR-mediated miniature currents (minis) were recorded at –60 mV in ACSF containing 4 m m CaCl2 , 2 m m MgCl2 , and 1 μ m TTX (Calbiochem).

Lysis:

Article Title: The Mechanoenzymatic Core of Dynamin-related Protein 1 Comprises the Minimal Machinery Required for Membrane Constriction *
Article Snippet: .. The overnight culture was harvested, and cell pellets were resuspended in a cell lysis/binding buffer (500 m m arginine, pH 8, 300 m m NaCl, 10 m m β-mercaptoethanol (BME), 5.0 m m magnesium chloride, 1.0 m m imidazole, and 2 m m CaCl2 ) with 100 ng/μl lysosome (Sigma), 4 units/ml DNase (Sigma), and complete EDTA-free protease inhibitor mixture tablets (Roche). .. The resuspended cells were then passed through a micro fluidizer (M-110 Y; Microfluidics, Newton, MA), and the lysate was centrifuged in a Beckman Coulter Optima L-look Ultracentrifuge (50.2 Ti rotor) at 40,000 rpm (184,048 × g ) for 30 min at 4 °C.

Article Title: The Baseplate of Lactobacillus delbrueckii Bacteriophage Ld17 Harbors a Glycerophosphodiesterase *
Article Snippet: .. The bacterial pellet was thawed at room temperature, resuspended in lysis buffer (10 m m Tris-HCl (pH 7.5), 0.3 m NaCl, 10 m m CaCl2 ) supplemented with 30 mg/ml hen egg white lysozyme (Sigma-Aldrich), and incubated at 20 °C for 30 min. .. The bacterial suspension was subsequently placed on ice and disrupted with glass beads in a Mini Bead Beater (Biospec Products, Bartlesville, OK) for 90 s followed by 5 min on ice for three cycles.

Chick Chorioallantoic Membrane Assay:

Article Title: Calmodulin Wraps around Its Binding Domain in the Plasma Membrane Ca2+ Pump Anchored by a Novel 18-1 Motif *
Article Snippet: .. Upon addition of 5 m m CaCl2 , the cleared cell lysate was loaded onto a phenyl-Sepharose CL-4B column (Sigma), and CaM was eluted with 1 m m EGTA. .. The CaM fractions were further purified using a HiTrap Q column (Amersham Biosciences) and a 0–1 m NaCl gradient.

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  • 88
    Millipore tppe digestion buffer
    <t>TPPE</t> cleaves the reduced pro-BTH6 in the thionin domain. LC-ESI-MS analysis of the digestion of <t>carboxymethylated</t> myc-proBTH6-strep with recombinant TPPE. The acidic domain is cleaved several times with additional sites compared with the oxidized proprotein.
    Tppe Digestion Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tppe digestion buffer/product/Millipore
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    84
    Millipore acid 0 8 cacl2
    <t>TPPE</t> cleaves the reduced pro-BTH6 in the thionin domain. LC-ESI-MS analysis of the digestion of <t>carboxymethylated</t> myc-proBTH6-strep with recombinant TPPE. The acidic domain is cleaved several times with additional sites compared with the oxidized proprotein.
    Acid 0 8 Cacl2, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    acid 0 8 cacl2 - by Bioz Stars, 2020-04
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    99
    Millipore cacl2
    <t>TPPE</t> cleaves the reduced pro-BTH6 in the thionin domain. LC-ESI-MS analysis of the digestion of <t>carboxymethylated</t> myc-proBTH6-strep with recombinant TPPE. The acidic domain is cleaved several times with additional sites compared with the oxidized proprotein.
    Cacl2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cacl2/product/Millipore
    Average 99 stars, based on 50 article reviews
    Price from $9.99 to $1999.99
    cacl2 - by Bioz Stars, 2020-04
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      Buy from Supplier

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    TPPE cleaves the reduced pro-BTH6 in the thionin domain. LC-ESI-MS analysis of the digestion of carboxymethylated myc-proBTH6-strep with recombinant TPPE. The acidic domain is cleaved several times with additional sites compared with the oxidized proprotein.

    Journal: The Journal of Biological Chemistry

    Article Title: Isolation and Characterization of a Thionin Proprotein-processing Enzyme from Barley *

    doi: 10.1074/jbc.M115.647859

    Figure Lengend Snippet: TPPE cleaves the reduced pro-BTH6 in the thionin domain. LC-ESI-MS analysis of the digestion of carboxymethylated myc-proBTH6-strep with recombinant TPPE. The acidic domain is cleaved several times with additional sites compared with the oxidized proprotein.

    Article Snippet: The reduced and cysteine-carboxymethylated protein samples were dialyzed against TPPE digestion buffer (25 m m MES, 100 m m NaCl, 10 m m CaCl2 , pH 6.5) using Amicon Ultra 10K ultrafiltration centrifugal filters (Millipore) according to the recommendations of the manufacturer.

    Techniques: Mass Spectrometry, Recombinant