mgcl2  (Thermo Fisher)


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    Name:
    MgCl2 magnesium chloride
    Description:
    Thermo Scientific PCR reagents meet high quality standards to generate accurate results from even the most challenging molecular biology applications Magnesium ions Mg2 are an important component of PCR reactions where they interact with the DNA template dNTPs and Taq DNA Polymerase
    Catalog Number:
    ab0359
    Price:
    None
    Applications:
    PCR|PCR & Real-Time PCR|Routine PCR
    Category:
    Lab Reagents and Chemicals
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    Structured Review

    Thermo Fisher mgcl2
    <t>MgCl2:</t> Melt® results.
    Thermo Scientific PCR reagents meet high quality standards to generate accurate results from even the most challenging molecular biology applications Magnesium ions Mg2 are an important component of PCR reactions where they interact with the DNA template dNTPs and Taq DNA Polymerase
    https://www.bioz.com/result/mgcl2/product/Thermo Fisher
    Average 99 stars, based on 14263 article reviews
    Price from $9.99 to $1999.99
    mgcl2 - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "Rapid and simple comparison of messenger RNA levels using real-time PCR"

    Article Title: Rapid and simple comparison of messenger RNA levels using real-time PCR

    Journal: Biological Procedures Online

    doi: 10.1251/bpo114

    MgCl2: Melt® results.
    Figure Legend Snippet: MgCl2: Melt® results.

    Techniques Used:

    Optimization of MgCl2 concentration in samples.
    Figure Legend Snippet: Optimization of MgCl2 concentration in samples.

    Techniques Used: Concentration Assay

    2) Product Images from "Multi-pathogens sequence containing plasmids as positive controls for universal detection of potential agents of bioterrorism"

    Article Title: Multi-pathogens sequence containing plasmids as positive controls for universal detection of potential agents of bioterrorism

    Journal: BMC Microbiology

    doi: 10.1186/1471-2180-4-21

    Construction of DNA control plasmid designed for the 4 CDC Category A DNA agents (Smallpox virus [seq1], Bacillus anthracis [seq2], Francisella tularensis [seq3], and Yersinia pestis [seq4]). Assembling of the smallpox virus and B. anthracis sequences is presented as an example. Successive steps are indicated by framed numbers. 1, PCR amplification of the two matrix sequences by primers consisting of the stabilization and the restriction site sequences (italics). PCR reactions were carried out in a volume of 50 μl that included 10 mM Tris-HCl [pH 9.0], 1.5 mM MgCl2, 50 mM KCl, 0.1% Triton X-100, 200 μM each dNTP, 0.4 μM of each oligonucleotide primer, 0.4 μM of the single stranded DNA, and 1.5 U of Taq DNA polymerase (Invitrogen, Cergy-Pontoise, France). The thermocycler (Biometra, Göttingen, Germany) profile was 5 min at 95°C, followed by 35 cycles of 30 sec at 95°C, 30 sec at 55°C, and 1 min at 72°C, and terminated by a final extension for 7 min at 72°C. PCR products were electrophorezed in 3% TAE-agarose gel containing ethidium bromide and visualized under UV transillumination. Column purification of the PCR products. PCR products of the expected size were column-purified by using the QIAquick PCR Purification Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions, and eluted in 50 μl of RNase free distillated water. When two bands or more were observed by gel analysis, the band of expected size was excised from the gel and purified by glass milk extraction with the GenClean III Kit (Q-Bio-Gene, Carlsbad CA, USA). 2, assemblage was conducted by pair, seq1 with seq2 (resulting in seq1-2). Equal volumes (10 μl) of purified seq1- and seq2-dsDNA were incubated at 37°C in the presence of Sac I . Sac I site is located at the 3' and 5' ends of seq1 and seq2, respectively. 3, the reaction product was column purified using the protocol aforementioned to discard the 15-nt DNA fragments corresponding to the 5' and 3' ends to avoid their re-ligation to their respective complementary sequences at step 5. 4, Overnight incubation at 4°C in the presence of T4 DNA ligase. Ten μl of the reaction was incubated with T4 DNA ligase (Roche, Basel, Switzerland) according to the manufaturer's instructions. 5, PCR amplification by using the external primers (italics) was performed according to the protocol described at step 1. Then column purification using the protocol detailed at step 2 of the resulting PCR product. At this step the seq1-2 PCR product may be cloned into PGEM-T for storage. The same procedure was performed for seq3 and seq4. Ultimately, seq1-2 and seq3-4 were assembled by using the same protocol (sections 1–9). The final product cloned into PGEM-T plasmid includes seq1-2-3-4 flanked by the two Sseq and restriction sites.
    Figure Legend Snippet: Construction of DNA control plasmid designed for the 4 CDC Category A DNA agents (Smallpox virus [seq1], Bacillus anthracis [seq2], Francisella tularensis [seq3], and Yersinia pestis [seq4]). Assembling of the smallpox virus and B. anthracis sequences is presented as an example. Successive steps are indicated by framed numbers. 1, PCR amplification of the two matrix sequences by primers consisting of the stabilization and the restriction site sequences (italics). PCR reactions were carried out in a volume of 50 μl that included 10 mM Tris-HCl [pH 9.0], 1.5 mM MgCl2, 50 mM KCl, 0.1% Triton X-100, 200 μM each dNTP, 0.4 μM of each oligonucleotide primer, 0.4 μM of the single stranded DNA, and 1.5 U of Taq DNA polymerase (Invitrogen, Cergy-Pontoise, France). The thermocycler (Biometra, Göttingen, Germany) profile was 5 min at 95°C, followed by 35 cycles of 30 sec at 95°C, 30 sec at 55°C, and 1 min at 72°C, and terminated by a final extension for 7 min at 72°C. PCR products were electrophorezed in 3% TAE-agarose gel containing ethidium bromide and visualized under UV transillumination. Column purification of the PCR products. PCR products of the expected size were column-purified by using the QIAquick PCR Purification Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions, and eluted in 50 μl of RNase free distillated water. When two bands or more were observed by gel analysis, the band of expected size was excised from the gel and purified by glass milk extraction with the GenClean III Kit (Q-Bio-Gene, Carlsbad CA, USA). 2, assemblage was conducted by pair, seq1 with seq2 (resulting in seq1-2). Equal volumes (10 μl) of purified seq1- and seq2-dsDNA were incubated at 37°C in the presence of Sac I . Sac I site is located at the 3' and 5' ends of seq1 and seq2, respectively. 3, the reaction product was column purified using the protocol aforementioned to discard the 15-nt DNA fragments corresponding to the 5' and 3' ends to avoid their re-ligation to their respective complementary sequences at step 5. 4, Overnight incubation at 4°C in the presence of T4 DNA ligase. Ten μl of the reaction was incubated with T4 DNA ligase (Roche, Basel, Switzerland) according to the manufaturer's instructions. 5, PCR amplification by using the external primers (italics) was performed according to the protocol described at step 1. Then column purification using the protocol detailed at step 2 of the resulting PCR product. At this step the seq1-2 PCR product may be cloned into PGEM-T for storage. The same procedure was performed for seq3 and seq4. Ultimately, seq1-2 and seq3-4 were assembled by using the same protocol (sections 1–9). The final product cloned into PGEM-T plasmid includes seq1-2-3-4 flanked by the two Sseq and restriction sites.

    Techniques Used: Plasmid Preparation, Polymerase Chain Reaction, Amplification, Size-exclusion Chromatography, Agarose Gel Electrophoresis, Purification, Incubation, Ligation, Clone Assay

    3) Product Images from "A novel dengue virus detection method that couples DNAzyme and gold nanoparticle approaches"

    Article Title: A novel dengue virus detection method that couples DNAzyme and gold nanoparticle approaches

    Journal: Virology Journal

    doi: 10.1186/1743-422X-10-201

    Assessment of DDZ-AuNP specificity. A) DENV-2 and CHIKV (1 × 106/mL each) were placed in a buffered solution containing 10 mM MgCl2, 1 × 105 DDZ-AuNP particles/mL, 1.5 M NaCl, and 0.5% (w/v) SDS. Eppendorf tubes containing these mixes were incubated at 37°C for 30 minutes and photographed. CHIKV = chikungunya virus. DENV-2 = dengue virus serotype 2, DDZ-M = anti-dengue virus DNAzyme, DDZin-M = inactive anti-dengue virus DNAzyme. B) Analysis of DENV detection by UV/Vis Spectrophotometry. Samples were assembled as was performed for in A, mixed, incubated at 37°C for 5 minutes, and spectrophotometric analysis was performed using the ND-1000 spectrophotometer. C) Detection of DENV by DDZ-M-AuNP in comparison to several other flaviviruses. The specificity of our DDZ-M-AuNP device to detect DENV and not other fellow flavivirus members YFV, JEV or ZV was determined as described earlier (see Figure 4 ). D) An alignment was performed on consensus sequences of each of the four DENV serotypes to determine the most optimal regions for the design of serotype specific DDZ-AuNP devices by determining the region of least conservation one serotype has with the other DENV serotypes. E) Colorimetric serotype-specific detection of DENV. Cell culture supernatants from C6/36 cells mock infected (Mock), or from cells infected with either DENV serotypes 1 through 4 or CHIKV (1 × 106/mL each) were placed in buffered solutions containing the necessary cofactors and AuNPs tethered with our all purpose DENV serotype specific DNAzyme, DDZ-M, or one of the serotype-specific DDZs (designated DDZ-1 through-4) designed to specifically target the corresponding DENV serotype (Table 1 ). Eppendorf tubes containing these mixes were incubated at 37°C for 5 minutes and photographed.
    Figure Legend Snippet: Assessment of DDZ-AuNP specificity. A) DENV-2 and CHIKV (1 × 106/mL each) were placed in a buffered solution containing 10 mM MgCl2, 1 × 105 DDZ-AuNP particles/mL, 1.5 M NaCl, and 0.5% (w/v) SDS. Eppendorf tubes containing these mixes were incubated at 37°C for 30 minutes and photographed. CHIKV = chikungunya virus. DENV-2 = dengue virus serotype 2, DDZ-M = anti-dengue virus DNAzyme, DDZin-M = inactive anti-dengue virus DNAzyme. B) Analysis of DENV detection by UV/Vis Spectrophotometry. Samples were assembled as was performed for in A, mixed, incubated at 37°C for 5 minutes, and spectrophotometric analysis was performed using the ND-1000 spectrophotometer. C) Detection of DENV by DDZ-M-AuNP in comparison to several other flaviviruses. The specificity of our DDZ-M-AuNP device to detect DENV and not other fellow flavivirus members YFV, JEV or ZV was determined as described earlier (see Figure 4 ). D) An alignment was performed on consensus sequences of each of the four DENV serotypes to determine the most optimal regions for the design of serotype specific DDZ-AuNP devices by determining the region of least conservation one serotype has with the other DENV serotypes. E) Colorimetric serotype-specific detection of DENV. Cell culture supernatants from C6/36 cells mock infected (Mock), or from cells infected with either DENV serotypes 1 through 4 or CHIKV (1 × 106/mL each) were placed in buffered solutions containing the necessary cofactors and AuNPs tethered with our all purpose DENV serotype specific DNAzyme, DDZ-M, or one of the serotype-specific DDZs (designated DDZ-1 through-4) designed to specifically target the corresponding DENV serotype (Table 1 ). Eppendorf tubes containing these mixes were incubated at 37°C for 5 minutes and photographed.

    Techniques Used: Incubation, Spectrophotometry, Cell Culture, Infection

    4) Product Images from "Probing the Oligomeric Assemblies of Pea Porphobilinogen Synthase by Analytical Ultracentrifugation †"

    Article Title: Probing the Oligomeric Assemblies of Pea Porphobilinogen Synthase by Analytical Ultracentrifugation †

    Journal:

    doi: 10.1021/bi801128d

    Analysis of Pea PBGS in the presence of 10 mM MgCl2
    Figure Legend Snippet: Analysis of Pea PBGS in the presence of 10 mM MgCl2

    Techniques Used:

    Analysis of Pea PBGS in the presence of 10 mM MgCl2
    Figure Legend Snippet: Analysis of Pea PBGS in the presence of 10 mM MgCl2

    Techniques Used:

    5) Product Images from "Conformational Changes during Nucleotide Selection by Sulfolobus solfataricus DNA Polymerase Dpo4 *"

    Article Title: Conformational Changes during Nucleotide Selection by Sulfolobus solfataricus DNA Polymerase Dpo4 *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.009506

    Overview of Changes in HDX as a Function of Primer/Template DNA, MgCl2 , and Incoming dNTP
    Figure Legend Snippet: Overview of Changes in HDX as a Function of Primer/Template DNA, MgCl2 , and Incoming dNTP

    Techniques Used:

    6) Product Images from "Recognition of the small regulatory RNA RydC by the bacterial Hfq protein"

    Article Title: Recognition of the small regulatory RNA RydC by the bacterial Hfq protein

    Journal: eLife

    doi: 10.7554/eLife.05375

    Size exclusion chromatography dynamic light scattering (SEC-MALS). ( A ) Elution profile for the RydC:Hfq complex from a Superdex 200 column superimposed on the estimated molecular masses. Fractions over the peak were analysed with a denaturing protein gel ( B ) and a denaturing RNA gel ( C ), confirming that both RydC and Hfq are present in the peak. The table in the lower panel summarises the results of the SEC-MALS data evaluations. SEC-MALS measurements were made with a Superdex 200 10/300 column connected to a miniDAWN TREOS multi-angle static Light Scattering and an Optilab T-rEX (refractometer with EXtended range) detector (Wyatt Technology Corporation, Santa Barbara CA, USA). Mixtures of protein and RNA in different molar ratios were run through an S200 10/300 size exclusion chromatography column equilibrated in buffer A (50 mM Tris, pH 7.5, 50 mM NaCl, 50 mM KCl, 5 mM MgCl2, 2 mM DTT). Both RydC and Hfq were diluted in buffer A
    Figure Legend Snippet: Size exclusion chromatography dynamic light scattering (SEC-MALS). ( A ) Elution profile for the RydC:Hfq complex from a Superdex 200 column superimposed on the estimated molecular masses. Fractions over the peak were analysed with a denaturing protein gel ( B ) and a denaturing RNA gel ( C ), confirming that both RydC and Hfq are present in the peak. The table in the lower panel summarises the results of the SEC-MALS data evaluations. SEC-MALS measurements were made with a Superdex 200 10/300 column connected to a miniDAWN TREOS multi-angle static Light Scattering and an Optilab T-rEX (refractometer with EXtended range) detector (Wyatt Technology Corporation, Santa Barbara CA, USA). Mixtures of protein and RNA in different molar ratios were run through an S200 10/300 size exclusion chromatography column equilibrated in buffer A (50 mM Tris, pH 7.5, 50 mM NaCl, 50 mM KCl, 5 mM MgCl2, 2 mM DTT). Both RydC and Hfq were diluted in buffer A

    Techniques Used: Size-exclusion Chromatography

    Related Articles

    Amplification:

    Article Title: Identification of the Main Promoter Directing Cereulide Biosynthesis in Emetic Bacillus cereus and Its Application for Real-Time Monitoring of ces Gene Expression in Foods ▿ Gene Expression in Foods ▿ †
    Article Snippet: .. The 50-μl PCR mixture (10 ng DNA, 0.5 μM each primer, 1.5 mM MgCl2 , 0.4 mM each deoxynucleoside triphosphate [dNTP], 1.25 U ThermoStart Taq polymerase [all from ABgene]) was activated (95°C for 15 min), followed by 30 amplification cycles (95°C for 30 s, 60°C for 45 s, and 72°C for 1 min) and an elongation step (72°C for 5 min). .. Total DNA was isolated as described previously , and plasmid DNA was prepared using standard procedures.

    Spectrophotometry:

    Article Title: A novel dengue virus detection method that couples DNAzyme and gold nanoparticle approaches
    Article Snippet: .. The stability of DDZ-M-AuNP was tested against increasing concentrations of MgCl2 (0 mM to 20 mM) at room temperature every 5 minutes for up to 30 minutes (Figure ), and absorbencies were measured with a NanoDrop spectrophotometer at 520 nm. ..

    SYBR Green Assay:

    Article Title: Rapid and simple comparison of messenger RNA levels using real-time PCR
    Article Snippet: .. Each sample consisted of: 50 ng cDNA, 3 mM MgCl2 , 200 μM dNTP, 500 nM of primers, 2 μl of 10X PCR buffer, 0.1 unit of Taq polymerase and SYBR® Green (Molecular Probe, Eugene, OR; 1/30 000 dilution), in a reaction volume of 20 μl. ..

    Concentration Assay:

    Article Title: Association of COX-2 Promoter Polymorphisms -765G/C and -1195A/G with Migraine
    Article Snippet: .. Therefore, the final volume of 25 μl PCR reactions in 0.2 ml tubes containing 500 ng/μl of DNA template, 2 mM of MgCl2 concentration, 2 mM dNTPs, 10 pmol/μl of each primer, 5 μl of 10X PCR buffer and 1 U of Taq DNA polymerase (Fermentas, Germany) was performed. .. Thermal PCR conditions consisted of denaturation phase for 5 min at 95 °C, followed by 30 cycles of 94 °C for 1 min, temperature of 59 °C for COX-2-1195A>G (rs689466) primer and 56 °C for COX-2-765G>C (rs20417) primer for 1 min, and 72 °C for 1 min, with a final extension for 5 min at 72 °C.

    Incubation:

    Article Title: A unique insertion in STARD9's motor domain regulates its stability
    Article Snippet: .. After 3 h, a master mix containing all the ubiquitination components (ubiquitin [Enzo Life Sciences], ROC1, E1, E2, Skp1, with or without β-TrCP, with or without CUL1, in a buffer containing 20 mM HEPES, 5 mM NaCl, 5 mM MgCl2 , DTT, MG132, and protease and phosphatase inhibitor cocktail (Thermo Scientific) and ATP regeneration system was added to the tubes and further incubated for 90 min at 30°C. ..

    Polymerase Chain Reaction:

    Article Title: Identification of the Main Promoter Directing Cereulide Biosynthesis in Emetic Bacillus cereus and Its Application for Real-Time Monitoring of ces Gene Expression in Foods ▿ Gene Expression in Foods ▿ †
    Article Snippet: .. The 50-μl PCR mixture (10 ng DNA, 0.5 μM each primer, 1.5 mM MgCl2 , 0.4 mM each deoxynucleoside triphosphate [dNTP], 1.25 U ThermoStart Taq polymerase [all from ABgene]) was activated (95°C for 15 min), followed by 30 amplification cycles (95°C for 30 s, 60°C for 45 s, and 72°C for 1 min) and an elongation step (72°C for 5 min). .. Total DNA was isolated as described previously , and plasmid DNA was prepared using standard procedures.

    Article Title: Association of COX-2 Promoter Polymorphisms -765G/C and -1195A/G with Migraine
    Article Snippet: .. Therefore, the final volume of 25 μl PCR reactions in 0.2 ml tubes containing 500 ng/μl of DNA template, 2 mM of MgCl2 concentration, 2 mM dNTPs, 10 pmol/μl of each primer, 5 μl of 10X PCR buffer and 1 U of Taq DNA polymerase (Fermentas, Germany) was performed. .. Thermal PCR conditions consisted of denaturation phase for 5 min at 95 °C, followed by 30 cycles of 94 °C for 1 min, temperature of 59 °C for COX-2-1195A>G (rs689466) primer and 56 °C for COX-2-765G>C (rs20417) primer for 1 min, and 72 °C for 1 min, with a final extension for 5 min at 72 °C.

    Article Title: Rapid and simple comparison of messenger RNA levels using real-time PCR
    Article Snippet: .. Each sample consisted of: 50 ng cDNA, 3 mM MgCl2 , 200 μM dNTP, 500 nM of primers, 2 μl of 10X PCR buffer, 0.1 unit of Taq polymerase and SYBR® Green (Molecular Probe, Eugene, OR; 1/30 000 dilution), in a reaction volume of 20 μl. ..

    Article Title: Rapid, Quantitative PCR Monitoring of Growth of Clostridium botulinum Type E in Modified-Atmosphere-Packaged Fish
    Article Snippet: .. Reaction volumes (50 μl) for the PCR consisted of 5 μl of template DNA; 5.0 mM MgCl2 ; 5 μl of 10×TaqMan buffer A; 200 nM each primer; 200 μM each dATP, dCTP, and dGTP; 400 μM dUTP; 1.25 U of AmpliTaq Gold DNA polymerase (Applied Biosystems); 0.5 U of uracil- N -glycosidase (AmpErase UNG; Applied Biosystems); and 100 nM TaqMan probe. .. Amplification and detection were performed with the ABI PRISM 7700 sequence detector (Applied Biosystems).

    Article Title: Multi-pathogens sequence containing plasmids as positive controls for universal detection of potential agents of bioterrorism
    Article Snippet: .. PCR reactions were carried out in a volume of 50 μl that included 10 mM Tris-HCl [pH 9.0], 1.5 mM MgCl2, 50 mM KCl, 0.1% Triton X-100, 200 μM each dNTP, 0.4 μM of each oligonucleotide primer, 0.4 μM of the single stranded DNA, and 1.5 U of Taq DNA polymerase (Invitrogen, Cergy-Pontoise, France). .. The thermocycler (Biometra, Göttingen, Germany) profile was 5 min at 95°C, followed by 35 cycles of 30 sec at 95°C, 30 sec at 55°C, and 1 min at 72°C, and terminated by a final extension for 7 min at 72°C.

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  • 91
    Thermo Fisher cellular atp contents
    The OXPHOS pathway in porcine intestinal epithelial cells is activated by L. gasseri LA39. (A) KEGG pathway enrichment analysis for differentially expressed proteins. Values in the column indicate the normalized p -values (-log10). The most enriched KEGG pathway is displayed at the top of the column. (B) Differential expression profiles of proteins involved in OXPHOS metabolic pathway. All differentially expressed proteins are shown below the corresponding complex in OXPHOS pathway map. Protein marked with a red box showed a significantly increased expression comparing the L. gasseri LA39 group with the Ctrl group. (C) Western blotting analysis of the expression levels of TCIRG1, UQCRC2, and GAPDH in <t>IPEC-J2</t> cells. Normalization and quantitation of TCIRG1/GAPDH and UQCRC2/GAPDH are shown in the corresponding bar chart. (D,E) The relative mRNA expression of TCIRG1, UQCRC2, and GAPDH in IPEC-J2 cells. Normalization and quantitation of TCIRG1/GAPDH (D) and UQCRC2/GAPDH (E) were shown in corresponding bar chart. (F) <t>ATP</t> levels of IPEC-J2 cells. The ATP concentration (nmol/L) was normalized to the total protein concentration (mg/L) of WCLs. Data are shown as mean ± SEM; n = 3 (C); n = 5 (E); n = 6 (F). ∗∗ p
    Cellular Atp Contents, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cellular atp contents/product/Thermo Fisher
    Average 91 stars, based on 2 article reviews
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    93
    Thermo Fisher atp determination intracellular atp levels
    Mitochondrial dysfunction is observed in CD8 + but not <t>CD4</t> + EMRA T cell subsets. (a) Representative flow cytometry plots and cumulative graphs of TMRE staining from middle‐aged donors showing membrane potential in CD45RA/CD27 T cell subsets directly ex vivo defined showing the percentage of cortactin‐positive (a) CD4 + and (b) CD8 + T cells analysed directly ex vivo. Data expressed as mean ± SEM of six donors. (b) Mitochondrial ROS measured using MitoSOX by flow cytometry in CD4 + and CD8 + EMRA T cells from middle‐aged donors. Data expressed as mean ± SEM of six donors. (c) Mitochondrial ROS production expressed as a ratio of mitochondrial mass. Calculated from data shown in Figures 1 b and 2 . (d) γH2AX expression as determined by flow cytometry in CD45RA/CD27‐defined T cell subsets directly ex vivo from middle‐aged donors; the graph shows the mean ± SEM for five donors. (e) Oxygen consumption rates (OCR) of the EMRA CD4 + and CD8 + T cell subsets from middle‐aged donors were measured following a 15‐min stimulation with 0.5 µg/ml anti‐CD3 and 5 ng/ml IL‐2; the cells were then subjected to a metabolic stress test using the indicated mitochondrial inhibitors. Data are representative of four independent experiments. (f) The basal OCR, extracellular acidification rate (ECAR) and spare respiratory capacity were measured following a 15‐min stimulation with 0.5 µg/ml anti‐CD3 and 5 ng/ml IL‐2. Graphs show the mean ± SEM for four donors. (g) <t>ATP</t> concentration in EMRA T cell subsets from middle‐aged donors, graphs show the mean ± SEM for five donors. p ‐values were calculated using a t test. * p
    Atp Determination Intracellular Atp Levels, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
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    90
    Thermo Fisher pcr atp
    Global assessments of the myocardial bioenergetics of infarcted swine hearts changed in response to combined treatment with a fibrin patch and injected hiPSC-derived cardiac cells. (A) Global assessments of myocardial <t>ATP</t> metabolism were evaluated via an established 31 P MRS-MST experiment. The experiment consists of a control spectrum obtained without saturation ( a1 ) to quantify the magnetization (M0) of phosphocreatine <t>(PCr)</t> and ATPγ, a spectrum obtained with ATPγ saturated ( a2 ) to quantify the steady-state magnetization (Mss) of PCr, and a spectrum obtained with both Pi and PCr saturated ( a3 ) to quantify the steady-state magnetization of ATPγ; saturation was performed with BISTRO frequency-selective pulses applied at the positions indicated by the arrows in spectra a2 and a3 . This approach enables the rates of ATP utilization to be calculated in vivo, even when the resonance for inorganic phosphate (Pi) is obscured by the resonance of 2,3-diphosphoglycerate (2,3-DPG). (B-C) Spectra a1, a2, and a3 were used to calculate global measures of (B) the PCr/ATPγ ratio, and (C) the rate of the PCr→ATP reaction ( k PCR→ATP ) in the hearts of animals in the Cell+Patch, Patch, MI, and Sham groups. * P
    Pcr Atp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The OXPHOS pathway in porcine intestinal epithelial cells is activated by L. gasseri LA39. (A) KEGG pathway enrichment analysis for differentially expressed proteins. Values in the column indicate the normalized p -values (-log10). The most enriched KEGG pathway is displayed at the top of the column. (B) Differential expression profiles of proteins involved in OXPHOS metabolic pathway. All differentially expressed proteins are shown below the corresponding complex in OXPHOS pathway map. Protein marked with a red box showed a significantly increased expression comparing the L. gasseri LA39 group with the Ctrl group. (C) Western blotting analysis of the expression levels of TCIRG1, UQCRC2, and GAPDH in IPEC-J2 cells. Normalization and quantitation of TCIRG1/GAPDH and UQCRC2/GAPDH are shown in the corresponding bar chart. (D,E) The relative mRNA expression of TCIRG1, UQCRC2, and GAPDH in IPEC-J2 cells. Normalization and quantitation of TCIRG1/GAPDH (D) and UQCRC2/GAPDH (E) were shown in corresponding bar chart. (F) ATP levels of IPEC-J2 cells. The ATP concentration (nmol/L) was normalized to the total protein concentration (mg/L) of WCLs. Data are shown as mean ± SEM; n = 3 (C); n = 5 (E); n = 6 (F). ∗∗ p

    Journal: Frontiers in Microbiology

    Article Title: Lactobacillus gasseri LA39 Activates the Oxidative Phosphorylation Pathway in Porcine Intestinal Epithelial Cells

    doi: 10.3389/fmicb.2018.03025

    Figure Lengend Snippet: The OXPHOS pathway in porcine intestinal epithelial cells is activated by L. gasseri LA39. (A) KEGG pathway enrichment analysis for differentially expressed proteins. Values in the column indicate the normalized p -values (-log10). The most enriched KEGG pathway is displayed at the top of the column. (B) Differential expression profiles of proteins involved in OXPHOS metabolic pathway. All differentially expressed proteins are shown below the corresponding complex in OXPHOS pathway map. Protein marked with a red box showed a significantly increased expression comparing the L. gasseri LA39 group with the Ctrl group. (C) Western blotting analysis of the expression levels of TCIRG1, UQCRC2, and GAPDH in IPEC-J2 cells. Normalization and quantitation of TCIRG1/GAPDH and UQCRC2/GAPDH are shown in the corresponding bar chart. (D,E) The relative mRNA expression of TCIRG1, UQCRC2, and GAPDH in IPEC-J2 cells. Normalization and quantitation of TCIRG1/GAPDH (D) and UQCRC2/GAPDH (E) were shown in corresponding bar chart. (F) ATP levels of IPEC-J2 cells. The ATP concentration (nmol/L) was normalized to the total protein concentration (mg/L) of WCLs. Data are shown as mean ± SEM; n = 3 (C); n = 5 (E); n = 6 (F). ∗∗ p

    Article Snippet: Measurement of Cellular ATP Levels The total cellular ATP contents in IPEC-J2 cells and intestinal epithelial cells (including duodenum, jejunum, and ileum) from weaned piglets were measured by an ATP determination kit (Thermo Scientific, A22066) according to the manufacturer’s protocol.

    Techniques: Expressing, Western Blot, Quantitation Assay, Concentration Assay, Protein Concentration

    Fructose increases cellular ATP levels and the release of dopamine from the Neuro 2a cells. The cellular ATP contents (A) , levels of dopamine in culture medium (B) and cell viability (C) of the N2a cells determined at 72 hours after exposure to different concentrations of fructose (0, 12.5, 25, or 50 μM). Values are mean ± SEM of quadruplicate analyses. * P

    Journal: Journal of Biomedical Science

    Article Title: An increase in adenosine-5'-triphosphate (ATP) content in rostral ventrolateral medulla is engaged in the high fructose diet-induced hypertension

    doi: 10.1186/1423-0127-21-8

    Figure Lengend Snippet: Fructose increases cellular ATP levels and the release of dopamine from the Neuro 2a cells. The cellular ATP contents (A) , levels of dopamine in culture medium (B) and cell viability (C) of the N2a cells determined at 72 hours after exposure to different concentrations of fructose (0, 12.5, 25, or 50 μM). Values are mean ± SEM of quadruplicate analyses. * P

    Article Snippet: To determine cellular ATP level in the N2a cells, samples were collected in a lysis buffer (Thermo Fisher Scientific Inc.) and centrifuged at 13500 rpm for 15 min. Supernatant (10 μl) was incubated with ATP reaction mixture for 30 min and detected by the same procedures.

    Techniques:

    KHK gene knockdown prevents the fructose-induced increases in cellular pyruvate and ATP level as well as dopamine release from the N2a cells. Representative immunofluorescence images showing distribution of KHK in the N2a cells (A) on day 7 after the lentiviral transfection of small hairpin RNA of KHK (KHK shRNA) or control shRNA; as well as changes in KHK mRNA (B) , cellular pyruvate (C) and ATP contents (D) , and dopamine levels in the culture medium (E) , detected at 72 hours after the administration of different concentrations of fructose (0, 12.5, 25, or 50 μM) to the culture medium of the KHK shRNA or control shRNA-transfected N2a cells. Values are mean ± SEM of quadruplicate analyses. * P

    Journal: Journal of Biomedical Science

    Article Title: An increase in adenosine-5'-triphosphate (ATP) content in rostral ventrolateral medulla is engaged in the high fructose diet-induced hypertension

    doi: 10.1186/1423-0127-21-8

    Figure Lengend Snippet: KHK gene knockdown prevents the fructose-induced increases in cellular pyruvate and ATP level as well as dopamine release from the N2a cells. Representative immunofluorescence images showing distribution of KHK in the N2a cells (A) on day 7 after the lentiviral transfection of small hairpin RNA of KHK (KHK shRNA) or control shRNA; as well as changes in KHK mRNA (B) , cellular pyruvate (C) and ATP contents (D) , and dopamine levels in the culture medium (E) , detected at 72 hours after the administration of different concentrations of fructose (0, 12.5, 25, or 50 μM) to the culture medium of the KHK shRNA or control shRNA-transfected N2a cells. Values are mean ± SEM of quadruplicate analyses. * P

    Article Snippet: To determine cellular ATP level in the N2a cells, samples were collected in a lysis buffer (Thermo Fisher Scientific Inc.) and centrifuged at 13500 rpm for 15 min. Supernatant (10 μl) was incubated with ATP reaction mixture for 30 min and detected by the same procedures.

    Techniques: Immunofluorescence, Transfection, shRNA

    Mitochondrial dysfunction is observed in CD8 + but not CD4 + EMRA T cell subsets. (a) Representative flow cytometry plots and cumulative graphs of TMRE staining from middle‐aged donors showing membrane potential in CD45RA/CD27 T cell subsets directly ex vivo defined showing the percentage of cortactin‐positive (a) CD4 + and (b) CD8 + T cells analysed directly ex vivo. Data expressed as mean ± SEM of six donors. (b) Mitochondrial ROS measured using MitoSOX by flow cytometry in CD4 + and CD8 + EMRA T cells from middle‐aged donors. Data expressed as mean ± SEM of six donors. (c) Mitochondrial ROS production expressed as a ratio of mitochondrial mass. Calculated from data shown in Figures 1 b and 2 . (d) γH2AX expression as determined by flow cytometry in CD45RA/CD27‐defined T cell subsets directly ex vivo from middle‐aged donors; the graph shows the mean ± SEM for five donors. (e) Oxygen consumption rates (OCR) of the EMRA CD4 + and CD8 + T cell subsets from middle‐aged donors were measured following a 15‐min stimulation with 0.5 µg/ml anti‐CD3 and 5 ng/ml IL‐2; the cells were then subjected to a metabolic stress test using the indicated mitochondrial inhibitors. Data are representative of four independent experiments. (f) The basal OCR, extracellular acidification rate (ECAR) and spare respiratory capacity were measured following a 15‐min stimulation with 0.5 µg/ml anti‐CD3 and 5 ng/ml IL‐2. Graphs show the mean ± SEM for four donors. (g) ATP concentration in EMRA T cell subsets from middle‐aged donors, graphs show the mean ± SEM for five donors. p ‐values were calculated using a t test. * p

    Journal: Aging Cell

    Article Title: Mitochondrial mass governs the extent of human T cell senescence. Mitochondrial mass governs the extent of human T cell senescence

    doi: 10.1111/acel.13067

    Figure Lengend Snippet: Mitochondrial dysfunction is observed in CD8 + but not CD4 + EMRA T cell subsets. (a) Representative flow cytometry plots and cumulative graphs of TMRE staining from middle‐aged donors showing membrane potential in CD45RA/CD27 T cell subsets directly ex vivo defined showing the percentage of cortactin‐positive (a) CD4 + and (b) CD8 + T cells analysed directly ex vivo. Data expressed as mean ± SEM of six donors. (b) Mitochondrial ROS measured using MitoSOX by flow cytometry in CD4 + and CD8 + EMRA T cells from middle‐aged donors. Data expressed as mean ± SEM of six donors. (c) Mitochondrial ROS production expressed as a ratio of mitochondrial mass. Calculated from data shown in Figures 1 b and 2 . (d) γH2AX expression as determined by flow cytometry in CD45RA/CD27‐defined T cell subsets directly ex vivo from middle‐aged donors; the graph shows the mean ± SEM for five donors. (e) Oxygen consumption rates (OCR) of the EMRA CD4 + and CD8 + T cell subsets from middle‐aged donors were measured following a 15‐min stimulation with 0.5 µg/ml anti‐CD3 and 5 ng/ml IL‐2; the cells were then subjected to a metabolic stress test using the indicated mitochondrial inhibitors. Data are representative of four independent experiments. (f) The basal OCR, extracellular acidification rate (ECAR) and spare respiratory capacity were measured following a 15‐min stimulation with 0.5 µg/ml anti‐CD3 and 5 ng/ml IL‐2. Graphs show the mean ± SEM for four donors. (g) ATP concentration in EMRA T cell subsets from middle‐aged donors, graphs show the mean ± SEM for five donors. p ‐values were calculated using a t test. * p

    Article Snippet: 4.7 ATP determination Intracellular ATP levels were measured ex vivo on sorted CD4+ and CD8+ T cell subsets via a bioluminescence assay according to the manufacturer's instructions (Thermo Fisher).

    Techniques: Flow Cytometry, Staining, Ex Vivo, Expressing, Concentration Assay

    Global assessments of the myocardial bioenergetics of infarcted swine hearts changed in response to combined treatment with a fibrin patch and injected hiPSC-derived cardiac cells. (A) Global assessments of myocardial ATP metabolism were evaluated via an established 31 P MRS-MST experiment. The experiment consists of a control spectrum obtained without saturation ( a1 ) to quantify the magnetization (M0) of phosphocreatine (PCr) and ATPγ, a spectrum obtained with ATPγ saturated ( a2 ) to quantify the steady-state magnetization (Mss) of PCr, and a spectrum obtained with both Pi and PCr saturated ( a3 ) to quantify the steady-state magnetization of ATPγ; saturation was performed with BISTRO frequency-selective pulses applied at the positions indicated by the arrows in spectra a2 and a3 . This approach enables the rates of ATP utilization to be calculated in vivo, even when the resonance for inorganic phosphate (Pi) is obscured by the resonance of 2,3-diphosphoglycerate (2,3-DPG). (B-C) Spectra a1, a2, and a3 were used to calculate global measures of (B) the PCr/ATPγ ratio, and (C) the rate of the PCr→ATP reaction ( k PCR→ATP ) in the hearts of animals in the Cell+Patch, Patch, MI, and Sham groups. * P

    Journal: PLoS ONE

    Article Title: 31P NMR 2D Mapping of Creatine Kinase Forward Flux Rate in Hearts with Postinfarction Left Ventricular Remodeling in Response to Cell Therapy

    doi: 10.1371/journal.pone.0162149

    Figure Lengend Snippet: Global assessments of the myocardial bioenergetics of infarcted swine hearts changed in response to combined treatment with a fibrin patch and injected hiPSC-derived cardiac cells. (A) Global assessments of myocardial ATP metabolism were evaluated via an established 31 P MRS-MST experiment. The experiment consists of a control spectrum obtained without saturation ( a1 ) to quantify the magnetization (M0) of phosphocreatine (PCr) and ATPγ, a spectrum obtained with ATPγ saturated ( a2 ) to quantify the steady-state magnetization (Mss) of PCr, and a spectrum obtained with both Pi and PCr saturated ( a3 ) to quantify the steady-state magnetization of ATPγ; saturation was performed with BISTRO frequency-selective pulses applied at the positions indicated by the arrows in spectra a2 and a3 . This approach enables the rates of ATP utilization to be calculated in vivo, even when the resonance for inorganic phosphate (Pi) is obscured by the resonance of 2,3-diphosphoglycerate (2,3-DPG). (B-C) Spectra a1, a2, and a3 were used to calculate global measures of (B) the PCr/ATPγ ratio, and (C) the rate of the PCr→ATP reaction ( k PCR→ATP ) in the hearts of animals in the Cell+Patch, Patch, MI, and Sham groups. * P

    Article Snippet: PCr values obtained from the PCr/ATP of the spectra and chemically determined ATP levels by using a fluorometric ATP Assay Kit (ThermoFisher Scientific, USA) [ ].

    Techniques: Injection, Derivative Assay, Microscale Thermophoresis, Polymerase Chain Reaction, In Vivo

    Spatially localized assessments indicate that the myocardial bioenergetics of swine hearts are disrupted at the border zone of infarction and partially restored by combined treatment with a fibrin patch and injected hiPSC-derived cardiac cells. (A) Spatially localized assessments of myocardial energetics were performed via 31 P 2D-CSI MRS. Spectra were obtained both with (right) and without (left) saturation of the ATPγ resonance and used to calculate (B) the PCr/ATPγ ratios and (C) the rate of the PCr→ATP reaction ( k PCR→ATP ) at the border-zone of infarction and in remote (i.e., noninfarcted) myocardial regions from the hearts of animals in the Cell+Patch and Patch groups. (D) The Flux for the PCr→ATP reaction (Flux PCr→ATP ), as the product of multiplying k PCR→ATP , PCr/ATP ratios, and the ATP level that measured by chemical method. *, P

    Journal: PLoS ONE

    Article Title: 31P NMR 2D Mapping of Creatine Kinase Forward Flux Rate in Hearts with Postinfarction Left Ventricular Remodeling in Response to Cell Therapy

    doi: 10.1371/journal.pone.0162149

    Figure Lengend Snippet: Spatially localized assessments indicate that the myocardial bioenergetics of swine hearts are disrupted at the border zone of infarction and partially restored by combined treatment with a fibrin patch and injected hiPSC-derived cardiac cells. (A) Spatially localized assessments of myocardial energetics were performed via 31 P 2D-CSI MRS. Spectra were obtained both with (right) and without (left) saturation of the ATPγ resonance and used to calculate (B) the PCr/ATPγ ratios and (C) the rate of the PCr→ATP reaction ( k PCR→ATP ) at the border-zone of infarction and in remote (i.e., noninfarcted) myocardial regions from the hearts of animals in the Cell+Patch and Patch groups. (D) The Flux for the PCr→ATP reaction (Flux PCr→ATP ), as the product of multiplying k PCR→ATP , PCr/ATP ratios, and the ATP level that measured by chemical method. *, P

    Article Snippet: PCr values obtained from the PCr/ATP of the spectra and chemically determined ATP levels by using a fluorometric ATP Assay Kit (ThermoFisher Scientific, USA) [ ].

    Techniques: Injection, Derivative Assay, Polymerase Chain Reaction