Structured Review

Qiagen mgcl2
Mgcl2, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 1635 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mgcl2/product/Qiagen
Average 95 stars, based on 1635 article reviews
Price from $9.99 to $1999.99
mgcl2 - by Bioz Stars, 2020-07
95/100 stars

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Polymerase Chain Reaction:

Article Title: Identification of novel mutations of Insulin Receptor Substrate 1 (IRS1) in tumor samples of non-small cell lung cancer (NSCLC): Implications for aberrant insulin signaling in development of cancer
Article Snippet: .. PCR was performed in a total volume of 25 μL, containing 1x Qiagen Taq polymerase buffer (Qiagen, Germany), 2 mM MgCl2 , 6 mM dNTPs, 0.5 μM of each primer, 0.2 units Qiagen Taq DNA polymerase and 50 ng genomic DNA. .. PCR conditions were 5 min at 94 °C, followed by 35 cycles of 94 °C for 30 s, 58 °C for 1 min, 72 °C for 45 s, and one step of 72 °C for 10 min. PCR products were purified using a PCR Purification Kit (Invitrogen Carlsbad, CA), and the Big dye-terminator sequencing kit (Applied Biosystems, Foster City, CA) was used during amplification.

Article Title: Multiple-Locus Variable-Number Tandem-Repeat Analysis for Longitudinal Survey of Sources of Pseudomonas aeruginosa Infection in Cystic Fibrosis Patients ▿ Infection in Cystic Fibrosis Patients ▿ †
Article Snippet: .. PCRs were performed in reaction mixtures of 15 μl containing 5 to 10 ng of DNA, 1× PCR buffer, 1.5 mM MgCl2 , 1 U of Taq DNA polymerase (QIAGEN), 200 μM each deoxynucleoside triphosphate, and 0.3 μM each flanking primer (Eurogentec, Angers, France) in the presence of 0.5 M betain, as described previously ( ). .. Amplification was performed with a PTC 200 thermocycler (Bio-Rad, Marnes-la-Coquette, France) under the following conditions: an initial denaturation cycle for 5 min at 94°C and 35 cycles of denaturation for 30 s at 94°C, annealing for 30 s at 60°C, and elongation for 45 s at 72°C, plus a final elongation step for 10 min at 72°C.

Article Title: Distribution of Leptospira interrogans by Multispacer Sequence Typing in Urban Norway Rats (Rattus norvegicus): A Survey in France in 2011-2013
Article Snippet: .. PCR amplifications were performed in a final volume of 50 μl containing 35 μl of H2 O, 5 μl of 10× buffer (Qiagen), 2 μl of 25 mM MgCl2 , 1 μl of 10 mM dNTPs (Qiagen), 1 μl of forward primer (10 μM), 1 μl of reverse primer (10 μM), 0.3 μl of HotStarTaq DNA Polymerase (Qiagen) and 5 μl of target DNA. ..

Article Title: Fetal microtia and FGFR2 polymorphism
Article Snippet: .. The PCR system was 20 µl in total: A total of 50 ng genomic DNA, 1X PCR buffer, 3 mmol/l MgCl2 , 4 mmol/l dNTPs, 5 pmol/l primers and 1 unit TaqDNA polymerase (Qiagen, Inc., Valencia, CA, USA), and it was loaded on a CFX96 instrument for amplification (Bio-Rad Laboratories, Inc., Hercules, CA, USA). .. Parameters of qPCR amplification: 95°C for 30 sec, 58°C for 30 sec, 72°C for 30 sec, and then extension at 72°C for 7 min after 30 cycles.

Article Title: Molecular characterization and antimicrobial resistance profile of fecal contaminants and spoilage bacteria that emerge in rainbow trout (Oncorhynchus mykiss) farms
Article Snippet: .. Polymerase chain reaction (PCR) was performed in a total volume of 25 µl containing 2.5 µl 10x PCR Buffer, 5 µl 5x Q-Solution, 0.5 µl 0.2 mM dNTP mix (10 mM of each), 25 pmol of each gyrA primer, 37.5 pmol of each gyrB primer, 1 µl 25 mM MgCl2, 0.125 µl HotStarTaq DNA Polymerase (Qiagen, Venlo, Netherlands), 10 ng DNA template, and DNase⁄RNase-Free Distilled Water. .. The PCR reactions were conducted according to previously used methods [ , ].

Article Title: Identification of MHCII variants associated with chlamydial disease in the koala (Phascolarctos cinereus)
Article Snippet: .. The PCR amplifications were carried out in 25 µl reactions with 10–30 ng of DNA, 0.32 µM each primer (Sigma-Aldrich, Sydney, Australia), 1 × HotStarTaq DNA Polymerase PCR Buffer, 1 mM MgCl2 , 0.2 mM dNTPs, and 0.5 units of HotStarTaq DNA Polymerase (Qiagen). .. Cycle conditions were: initial activation at 95 °C for 15 min, followed by 35 cycles of 40 s denaturation at 95 °C, 40 s annealing at 57 °C or 54 °C (for DAB and DBB, respectively), and 45 s extension at 72 °C, and a final extension at 72 °C for 10 min. For OSCP analysis, the forward strand of PCR products was digested with Lambda exonuclease (New England Biolabs, Ipswich, MA, USA) at 37 °C for 45 min, and the reverse strand was subjected to electrophoresis in a 5% or 10% acrylamide gel (DAB and DBB, respectively) at 30 W for 5 h at 4 °C ( ).

Nested PCR:

Article Title: High prevalence of sexual Chlamydia trachomatis infection in young women from Marajó Island, in the Brazilian Amazon
Article Snippet: .. The first step of the nested PCR was run in a 0.5 μL volume containing 20 pmol/μL of each primer P1(B) and OMP2 and 5.0 μL of the DNA extracted from the endocervical secretion, 14 μL of sterile water, 1.0 μL of MgCl2 , 1.0 μL deoxynucleoside triphosphate (10mM), 2.5 μL of 10x buffer, and 0.5 μL of Hotstar Taq DNA Polymerase 1.5U (Qiagen). ..

Amplification:

Article Title: Fetal microtia and FGFR2 polymorphism
Article Snippet: .. The PCR system was 20 µl in total: A total of 50 ng genomic DNA, 1X PCR buffer, 3 mmol/l MgCl2 , 4 mmol/l dNTPs, 5 pmol/l primers and 1 unit TaqDNA polymerase (Qiagen, Inc., Valencia, CA, USA), and it was loaded on a CFX96 instrument for amplification (Bio-Rad Laboratories, Inc., Hercules, CA, USA). .. Parameters of qPCR amplification: 95°C for 30 sec, 58°C for 30 sec, 72°C for 30 sec, and then extension at 72°C for 7 min after 30 cycles.

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  • 85
    Qiagen atp binding protein
    The <t>IVa2</t> protein binds <t>ATP.</t> (A) UV cross-linking experiment with [α- 32 P]ATP and purified, wild-type IVa2-Strep protein either in the absence (−) or presence (+) of MgCl 2 , as indicated in lanes 1 and 2. Following cross-linking, samples were analyzed by SDS-polyacrylamide gel electrophoresis and the gel was exposed to a phosphorscreen. (B) Coomassie blue-stained SDS-polyacrylamide gel of UV cross-linked samples. Lane 1, molecular mass markers with indicated sizes in kilodaltons; lanes 2 and 3, wild-type IVa2-Strep protein (WT); lanes 4 and 5, A motif mutant IVa2-Strep protein (A Box); lanes 6 and 7, B motif mutant IVa2-Strep protein (B Box). Lanes 2, 4, and 6 are from reactions done in the presence of MgCl 2 , and lanes 3, 5, and 7 are from reactions done in the absence of MgCl 2 . (C) Radiolabeled image from the UV cross-linking experiment of wild-type and mutant IVa2-Strep proteins either in the absence (−) or presence (+) of MgCl 2 , as described for panel B.
    Atp Binding Protein, supplied by Qiagen, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 1 article reviews
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    92
    Qiagen atp competitive mtor inhibitor treated cells
    INK128, but not allosteric <t>mTOR</t> inhibitors, show enhanced activity in SNU423 and SNU449 HCC cells The HCC cell lines SNU423, SNU449 and Huh7 were treated with the allosteric mTOR inhibitors ( A ) rapamycin, ( B ) everolimus or ( C ) the <t>ATP-competitive</t> mTOR inhibitor INK128 for 48 h. Cell proliferation was determined by a WST-1 assay. ( D ) SNU423 cells were treated with vehicle control or 500 nM everolimus or INK128. Dashed lines represent the starting point of the in vitro migration assay at time 0. At 30 h after cell migration, the gap open areas were photographed and measured ( E ). * p
    Atp Competitive Mtor Inhibitor Treated Cells, supplied by Qiagen, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Qiagen steady state atp levels
    Basal ROS, antioxidant expression, <t>mtDNA</t> copy number and <t>ATP</t> levels Alstonville larvae had an advantage when fed the 1:2 P:C diet as the V161L ND4 amino acid change in complex I of Dahomey reduced the efficiency of ATP production. ( A ) Measurement of basal ROS shows higher levels in Dahomey fed the 1:2 P:C diet. ROS levels were similar when larvae were fed the 1:16 P:C diet (n = 9 biological rep/mitotype on the 1:2 P:C diet, and 8 biological rep/mitotype on the 1:16 P:C diet with 2 failed reactions in Alstonville). (B ). GstE1 and GstE5 expression was highest in Dahomey larvae fed the 1:2 P:C diet (n = 6 biological rep/mitotype/diet for both genes). ( C) Alstonville larvae had higher mtDNA copy number when fed the 1:2 P:C diet but both mitotypes had equivalent and lower copy number when fed the 1:16 P:C diet. MtDNA copy number show the relative expression of ND4 (ND4/Actin) and lrRNA (lrRNA/Rp49) (n = 8 biological rep/mitotype/diet with 2 failed reactions for ND4/Actin and 7 biological reps/mitotype/diet for lrRNA/Rp49 with 2 failed reactions). (D) Total cellular ATP levels were higher in Alstonville larvae fed the 1:2 P:C diet but were similar when fed the 1:16 P:C diet suggesting a mitohormetic response (n = 8 biological rep/mitotype/diet, with two failed reactions) Bars show mean± s.e.m. t-tests between mitotypes * p
    Steady State Atp Levels, supplied by Qiagen, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Qiagen γ 32 p atp
    Identification of the [ 32 P]P-Ser-HPr dephosphorylation product. The reaction products of HprK/P-catalyzed [ 32 P]P-Ser-HPr dephosphorylation were separated by TLC using 0.3 M KH 2 PO 4 as solvent. Radioactive standards for pyrophosphate and P-Ser-HPr (lanes 1 and 4, respectively); 1-[ 32 P]FBP formed from fructose-6-P and either [ 32 P]pyrophosphate or [γ- 32 <t>P]ATP</t> (lanes 2 and 3, respectively). Dephosphorylation of [ 32 P]P-Ser-HPr with B. subtilis HprK/P was carried out in the presence of fructose-6-P (lane 5), pyrophosphate-dependent phosphofructokinase (lane 6), and fructose-6-P and pyrophosphate-dependent phosphofructokinase (lane 7).
    γ 32 P Atp, supplied by Qiagen, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The IVa2 protein binds ATP. (A) UV cross-linking experiment with [α- 32 P]ATP and purified, wild-type IVa2-Strep protein either in the absence (−) or presence (+) of MgCl 2 , as indicated in lanes 1 and 2. Following cross-linking, samples were analyzed by SDS-polyacrylamide gel electrophoresis and the gel was exposed to a phosphorscreen. (B) Coomassie blue-stained SDS-polyacrylamide gel of UV cross-linked samples. Lane 1, molecular mass markers with indicated sizes in kilodaltons; lanes 2 and 3, wild-type IVa2-Strep protein (WT); lanes 4 and 5, A motif mutant IVa2-Strep protein (A Box); lanes 6 and 7, B motif mutant IVa2-Strep protein (B Box). Lanes 2, 4, and 6 are from reactions done in the presence of MgCl 2 , and lanes 3, 5, and 7 are from reactions done in the absence of MgCl 2 . (C) Radiolabeled image from the UV cross-linking experiment of wild-type and mutant IVa2-Strep proteins either in the absence (−) or presence (+) of MgCl 2 , as described for panel B.

    Journal: Journal of Virology

    Article Title: Adenovirus IVa2 Protein Binds ATP ▿

    doi: 10.1128/JVI.00882-08

    Figure Lengend Snippet: The IVa2 protein binds ATP. (A) UV cross-linking experiment with [α- 32 P]ATP and purified, wild-type IVa2-Strep protein either in the absence (−) or presence (+) of MgCl 2 , as indicated in lanes 1 and 2. Following cross-linking, samples were analyzed by SDS-polyacrylamide gel electrophoresis and the gel was exposed to a phosphorscreen. (B) Coomassie blue-stained SDS-polyacrylamide gel of UV cross-linked samples. Lane 1, molecular mass markers with indicated sizes in kilodaltons; lanes 2 and 3, wild-type IVa2-Strep protein (WT); lanes 4 and 5, A motif mutant IVa2-Strep protein (A Box); lanes 6 and 7, B motif mutant IVa2-Strep protein (B Box). Lanes 2, 4, and 6 are from reactions done in the presence of MgCl 2 , and lanes 3, 5, and 7 are from reactions done in the absence of MgCl 2 . (C) Radiolabeled image from the UV cross-linking experiment of wild-type and mutant IVa2-Strep proteins either in the absence (−) or presence (+) of MgCl 2 , as described for panel B.

    Article Snippet: These results demonstrate the importance of these amino acids in the Walker A and B motifs of IVa2 and implicate IVa2 as an ATP binding protein and a potential ATPase., P-labeled DNA probe containing the A1/A2 packaging repeats of Ad5 ( IVa2-Strep-WT protein was purified in one step from NP-40 lysates (100 mM Tris-HCl [pH 8.0], 300 mM NaCl, 0.5 mM ditiothreitol, 1% NP-40) of Sf9 cells by affinity chromatography with Strep-Tactin resin (Qiagen) according to the manufacturer's protocol.

    Techniques: Purification, Polyacrylamide Gel Electrophoresis, Staining, Mutagenesis

    INK128, but not allosteric mTOR inhibitors, show enhanced activity in SNU423 and SNU449 HCC cells The HCC cell lines SNU423, SNU449 and Huh7 were treated with the allosteric mTOR inhibitors ( A ) rapamycin, ( B ) everolimus or ( C ) the ATP-competitive mTOR inhibitor INK128 for 48 h. Cell proliferation was determined by a WST-1 assay. ( D ) SNU423 cells were treated with vehicle control or 500 nM everolimus or INK128. Dashed lines represent the starting point of the in vitro migration assay at time 0. At 30 h after cell migration, the gap open areas were photographed and measured ( E ). * p

    Journal: Oncotarget

    Article Title: CD44 positive and sorafenib insensitive hepatocellular carcinomas respond to the ATP-competitive mTOR inhibitor INK128

    doi: 10.18632/oncotarget.25430

    Figure Lengend Snippet: INK128, but not allosteric mTOR inhibitors, show enhanced activity in SNU423 and SNU449 HCC cells The HCC cell lines SNU423, SNU449 and Huh7 were treated with the allosteric mTOR inhibitors ( A ) rapamycin, ( B ) everolimus or ( C ) the ATP-competitive mTOR inhibitor INK128 for 48 h. Cell proliferation was determined by a WST-1 assay. ( D ) SNU423 cells were treated with vehicle control or 500 nM everolimus or INK128. Dashed lines represent the starting point of the in vitro migration assay at time 0. At 30 h after cell migration, the gap open areas were photographed and measured ( E ). * p

    Article Snippet: RNA extraction and quantitative RT-PCR Total RNA was isolated from ATP-competitive mTOR inhibitor treated cells using miRNeasy® Mini kit (Qiagen). cDNA was synthesized according to the manufacturer's protocol (Invitrogen).

    Techniques: Activity Assay, WST-1 Assay, In Vitro, Migration

    Basal ROS, antioxidant expression, mtDNA copy number and ATP levels Alstonville larvae had an advantage when fed the 1:2 P:C diet as the V161L ND4 amino acid change in complex I of Dahomey reduced the efficiency of ATP production. ( A ) Measurement of basal ROS shows higher levels in Dahomey fed the 1:2 P:C diet. ROS levels were similar when larvae were fed the 1:16 P:C diet (n = 9 biological rep/mitotype on the 1:2 P:C diet, and 8 biological rep/mitotype on the 1:16 P:C diet with 2 failed reactions in Alstonville). (B ). GstE1 and GstE5 expression was highest in Dahomey larvae fed the 1:2 P:C diet (n = 6 biological rep/mitotype/diet for both genes). ( C) Alstonville larvae had higher mtDNA copy number when fed the 1:2 P:C diet but both mitotypes had equivalent and lower copy number when fed the 1:16 P:C diet. MtDNA copy number show the relative expression of ND4 (ND4/Actin) and lrRNA (lrRNA/Rp49) (n = 8 biological rep/mitotype/diet with 2 failed reactions for ND4/Actin and 7 biological reps/mitotype/diet for lrRNA/Rp49 with 2 failed reactions). (D) Total cellular ATP levels were higher in Alstonville larvae fed the 1:2 P:C diet but were similar when fed the 1:16 P:C diet suggesting a mitohormetic response (n = 8 biological rep/mitotype/diet, with two failed reactions) Bars show mean± s.e.m. t-tests between mitotypes * p

    Journal: PLoS Genetics

    Article Title: Genotype to phenotype: Diet-by-mitochondrial DNA haplotype interactions drive metabolic flexibility and organismal fitness

    doi: 10.1371/journal.pgen.1007735

    Figure Lengend Snippet: Basal ROS, antioxidant expression, mtDNA copy number and ATP levels Alstonville larvae had an advantage when fed the 1:2 P:C diet as the V161L ND4 amino acid change in complex I of Dahomey reduced the efficiency of ATP production. ( A ) Measurement of basal ROS shows higher levels in Dahomey fed the 1:2 P:C diet. ROS levels were similar when larvae were fed the 1:16 P:C diet (n = 9 biological rep/mitotype on the 1:2 P:C diet, and 8 biological rep/mitotype on the 1:16 P:C diet with 2 failed reactions in Alstonville). (B ). GstE1 and GstE5 expression was highest in Dahomey larvae fed the 1:2 P:C diet (n = 6 biological rep/mitotype/diet for both genes). ( C) Alstonville larvae had higher mtDNA copy number when fed the 1:2 P:C diet but both mitotypes had equivalent and lower copy number when fed the 1:16 P:C diet. MtDNA copy number show the relative expression of ND4 (ND4/Actin) and lrRNA (lrRNA/Rp49) (n = 8 biological rep/mitotype/diet with 2 failed reactions for ND4/Actin and 7 biological reps/mitotype/diet for lrRNA/Rp49 with 2 failed reactions). (D) Total cellular ATP levels were higher in Alstonville larvae fed the 1:2 P:C diet but were similar when fed the 1:16 P:C diet suggesting a mitohormetic response (n = 8 biological rep/mitotype/diet, with two failed reactions) Bars show mean± s.e.m. t-tests between mitotypes * p

    Article Snippet: MtDNA copy number and steady-state ATP levels To determine mtDNA copy number, total DNA was extracted using DNeasy Blood and Tissue Kit (Qiagen 69582).

    Techniques: Expressing

    Identification of the [ 32 P]P-Ser-HPr dephosphorylation product. The reaction products of HprK/P-catalyzed [ 32 P]P-Ser-HPr dephosphorylation were separated by TLC using 0.3 M KH 2 PO 4 as solvent. Radioactive standards for pyrophosphate and P-Ser-HPr (lanes 1 and 4, respectively); 1-[ 32 P]FBP formed from fructose-6-P and either [ 32 P]pyrophosphate or [γ- 32 P]ATP (lanes 2 and 3, respectively). Dephosphorylation of [ 32 P]P-Ser-HPr with B. subtilis HprK/P was carried out in the presence of fructose-6-P (lane 5), pyrophosphate-dependent phosphofructokinase (lane 6), and fructose-6-P and pyrophosphate-dependent phosphofructokinase (lane 7).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Pyrophosphate-producing protein dephosphorylation by HPr kinase/phosphorylase: A relic of early life?

    doi: 10.1073/pnas.212410399

    Figure Lengend Snippet: Identification of the [ 32 P]P-Ser-HPr dephosphorylation product. The reaction products of HprK/P-catalyzed [ 32 P]P-Ser-HPr dephosphorylation were separated by TLC using 0.3 M KH 2 PO 4 as solvent. Radioactive standards for pyrophosphate and P-Ser-HPr (lanes 1 and 4, respectively); 1-[ 32 P]FBP formed from fructose-6-P and either [ 32 P]pyrophosphate or [γ- 32 P]ATP (lanes 2 and 3, respectively). Dephosphorylation of [ 32 P]P-Ser-HPr with B. subtilis HprK/P was carried out in the presence of fructose-6-P (lane 5), pyrophosphate-dependent phosphofructokinase (lane 6), and fructose-6-P and pyrophosphate-dependent phosphofructokinase (lane 7).

    Article Snippet: HprK/P was subsequently heat-inactivated (10 min at 70°C) before [32 P]P-Ser-HPr was separated from [γ-32 P]ATP by chromatography on nickel-nitrilotriacetic acid (Qiagen, Chatsworth, CA) and PD-10 (Pharmacia) columns.

    Techniques: De-Phosphorylation Assay, Thin Layer Chromatography