supercoiled dna plasmid puc19  (Thermo Fisher)


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    Name:
    pUC19 DNA
    Description:
    Thermo Scientific pUC19 vector is a small high copy number E coli plasmid 2686 bp in length It contains identical multiple cloning site MCS as pUC18 vector except that it is arranged in opposite orientation Highlights• Purified by chromatography using proprietary patented technology• More than 90 in the supercoiled form• Isolated from E coli dam dcm • For pUC18 DNA sequence pUC19 DNA sequence sequence analysis and map creation see free online REviewer tool Applications• Cloning• Sequencing of insert DNA pUC18 DNA Preparation of DNA molecular weight standardspUC18 19 plasmid contents and usage notes pUC18 19 plasmids contain • The pMB1 replicon rep responsible for the replication of plasmid source plasmid pBR322 The high copy number of pUC plasmids is a result of the lack of the rop gene and a single point mutation in the replicon rep of pMB1 • The bla gene encoding beta lactamase confers resistance to ampicillin source plasmid pBR322 It differs from that of pBR322 by two point mutations• The region of E coli lac operon containing a CAP protein binding site promoter Plac lac repressor binding site and the 5 terminal part of the lacZ gene encoding the N terminal fragment of beta galactosidase source M13mp18 19 This fragment whose synthesis can be induced by IPTG is capable of intra allelic alpha complementation with a defective form of beta galactosidase encoded by the host mutation delta lacZ M15 In the presence of IPTG bacteria synthesize both fragments of the enzyme and form blue colonies on media with X Gal Insertion of DNA into the MCS located within the lacZ gene codons 6 7 of lacZ are replaced by MCS inactivates the N terminal fragment of beta galactosidase and abolishes alpha complementation Bacteria carrying recombinant plasmids therefore give rise to white coloniesThe map shows enzymes that cut pUC18 DNA once Enzymes produced by Thermo Scientific are shown in orange The coordinates refer to the position of the first nucleotide in each recognition sequence The exact positions of the genetic elements are shown on the map termination codons included The bla gene nucleotides 2486 2418 complementary strand code for a signal peptide The LacZ polypeptide corresponding to wt beta galactosidase and essential for blue white screening ends at nt position 236 compl strand Another 30 codons in the same reading frame are derived from pBR322 The indicated rep region is sufficient to promote replication DNA replication initiates at position 866 1 and proceeds in the direction indicated Plasmids carrying the pMB1 and ColE1 replicons are incompatible but they are fully compatible with those carrying the p15A replicon pACYC177 pACYC184 pMB1 derived plasmids can be amplified using chloramphenicol Related ProductspUC18 DNApUC57 DNA
    Catalog Number:
    sd0061
    Price:
    None
    Applications:
    Agarose Gel Electrophoresis|Cloning|DNA & RNA Purification & Analysis|Restriction Enzyme Cloning|Nucleic Acid Gel Electrophoresis & Blotting
    Category:
    DNA Vectors Clones Purified Nucleic Acids Libraries
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    Structured Review

    Thermo Fisher supercoiled dna plasmid puc19
    Infection of cells with live M. fermentans inhibits the <t>DNA</t> relaxation activity of Topo I. MCF7 (A,B) and U251 (C,D) cancer cells were infected with live M. fermentans ( M.f ) for various intervals at MOI of 10 3 CFU/cell. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis. ( A,C ) A representative picture (n = 4–5) of Topo I DNA-relaxation activity. ( B,D ) Quantification analysis of Topo I activity. Symbols: R and S are the relaxed and <t>supercoiled</t> form of the <t>pUC19</t> DNA, respectively; Topo- no protein added to the reaction mixture. t -test: * p
    Thermo Scientific pUC19 vector is a small high copy number E coli plasmid 2686 bp in length It contains identical multiple cloning site MCS as pUC18 vector except that it is arranged in opposite orientation Highlights• Purified by chromatography using proprietary patented technology• More than 90 in the supercoiled form• Isolated from E coli dam dcm • For pUC18 DNA sequence pUC19 DNA sequence sequence analysis and map creation see free online REviewer tool Applications• Cloning• Sequencing of insert DNA pUC18 DNA Preparation of DNA molecular weight standardspUC18 19 plasmid contents and usage notes pUC18 19 plasmids contain • The pMB1 replicon rep responsible for the replication of plasmid source plasmid pBR322 The high copy number of pUC plasmids is a result of the lack of the rop gene and a single point mutation in the replicon rep of pMB1 • The bla gene encoding beta lactamase confers resistance to ampicillin source plasmid pBR322 It differs from that of pBR322 by two point mutations• The region of E coli lac operon containing a CAP protein binding site promoter Plac lac repressor binding site and the 5 terminal part of the lacZ gene encoding the N terminal fragment of beta galactosidase source M13mp18 19 This fragment whose synthesis can be induced by IPTG is capable of intra allelic alpha complementation with a defective form of beta galactosidase encoded by the host mutation delta lacZ M15 In the presence of IPTG bacteria synthesize both fragments of the enzyme and form blue colonies on media with X Gal Insertion of DNA into the MCS located within the lacZ gene codons 6 7 of lacZ are replaced by MCS inactivates the N terminal fragment of beta galactosidase and abolishes alpha complementation Bacteria carrying recombinant plasmids therefore give rise to white coloniesThe map shows enzymes that cut pUC18 DNA once Enzymes produced by Thermo Scientific are shown in orange The coordinates refer to the position of the first nucleotide in each recognition sequence The exact positions of the genetic elements are shown on the map termination codons included The bla gene nucleotides 2486 2418 complementary strand code for a signal peptide The LacZ polypeptide corresponding to wt beta galactosidase and essential for blue white screening ends at nt position 236 compl strand Another 30 codons in the same reading frame are derived from pBR322 The indicated rep region is sufficient to promote replication DNA replication initiates at position 866 1 and proceeds in the direction indicated Plasmids carrying the pMB1 and ColE1 replicons are incompatible but they are fully compatible with those carrying the p15A replicon pACYC177 pACYC184 pMB1 derived plasmids can be amplified using chloramphenicol Related ProductspUC18 DNApUC57 DNA
    https://www.bioz.com/result/supercoiled dna plasmid puc19/product/Thermo Fisher
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    supercoiled dna plasmid puc19 - by Bioz Stars, 2020-09
    98/100 stars

    Images

    1) Product Images from "Mycoplasma fermentans Inhibits the Activity of Cellular DNA Topoisomerase I by Activation of PARP1 and Alters the Efficacy of Its Anti-Cancer Inhibitor"

    Article Title: Mycoplasma fermentans Inhibits the Activity of Cellular DNA Topoisomerase I by Activation of PARP1 and Alters the Efficacy of Its Anti-Cancer Inhibitor

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0072377

    Infection of cells with live M. fermentans inhibits the DNA relaxation activity of Topo I. MCF7 (A,B) and U251 (C,D) cancer cells were infected with live M. fermentans ( M.f ) for various intervals at MOI of 10 3 CFU/cell. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis. ( A,C ) A representative picture (n = 4–5) of Topo I DNA-relaxation activity. ( B,D ) Quantification analysis of Topo I activity. Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively; Topo- no protein added to the reaction mixture. t -test: * p
    Figure Legend Snippet: Infection of cells with live M. fermentans inhibits the DNA relaxation activity of Topo I. MCF7 (A,B) and U251 (C,D) cancer cells were infected with live M. fermentans ( M.f ) for various intervals at MOI of 10 3 CFU/cell. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis. ( A,C ) A representative picture (n = 4–5) of Topo I DNA-relaxation activity. ( B,D ) Quantification analysis of Topo I activity. Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively; Topo- no protein added to the reaction mixture. t -test: * p

    Techniques Used: Infection, Activity Assay, Agarose Gel Electrophoresis

    Sonicated M. fermentans Protein inhibits the DNA relaxation activity of Topo I. MCF7 (A–C) and U251 (D–F) cancer cells were treated for 24 hrs with various concentrations of M. fermentans proteins. Total nuclear proteins (12.5 ng) were added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis. ( A,D ) A representative picture (n = 4–5) of Topo I DNA relaxation activity. ( B,E ) Quantification analysis of Topo I activity. ( C,F ) Topo I protein level examined by Western blot analysis. Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture. t -test: * p
    Figure Legend Snippet: Sonicated M. fermentans Protein inhibits the DNA relaxation activity of Topo I. MCF7 (A–C) and U251 (D–F) cancer cells were treated for 24 hrs with various concentrations of M. fermentans proteins. Total nuclear proteins (12.5 ng) were added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis. ( A,D ) A representative picture (n = 4–5) of Topo I DNA relaxation activity. ( B,E ) Quantification analysis of Topo I activity. ( C,F ) Topo I protein level examined by Western blot analysis. Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture. t -test: * p

    Techniques Used: Sonication, Activity Assay, Agarose Gel Electrophoresis, Western Blot

    PARP inhibitor prevented the Mycoplasma-induced inhibitory effect on Topo I activity. MCF7 cells were pre-incubated with 3-aminobenzamide (3AB) for 1 hr at various concentrations followed by M. fermentans infection (MOI of 10 3 CFU/cell) for an additional 6 hrs. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis (A) and quantification of Topo I activity was performed (B). Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture t -test: * p
    Figure Legend Snippet: PARP inhibitor prevented the Mycoplasma-induced inhibitory effect on Topo I activity. MCF7 cells were pre-incubated with 3-aminobenzamide (3AB) for 1 hr at various concentrations followed by M. fermentans infection (MOI of 10 3 CFU/cell) for an additional 6 hrs. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis (A) and quantification of Topo I activity was performed (B). Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture t -test: * p

    Techniques Used: Activity Assay, Incubation, Infection, Agarose Gel Electrophoresis

    M. fermentans diminished the CPT inhibition effect on the DNA relaxation activity of Topo I. MCF7 cells were infected with M. fermentans ( M.f ) for 6 hrs followed by CPT (30 µM) treatments for additional 1.5 hrs. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis. ( A ) A representative picture n = 5, of Topo I DNA-relaxation activity. ( B ) Quantification analysis of Topo I activity. ( C ) Topo I protein level examined by Western blot analysis. Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture t -test: * p
    Figure Legend Snippet: M. fermentans diminished the CPT inhibition effect on the DNA relaxation activity of Topo I. MCF7 cells were infected with M. fermentans ( M.f ) for 6 hrs followed by CPT (30 µM) treatments for additional 1.5 hrs. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis. ( A ) A representative picture n = 5, of Topo I DNA-relaxation activity. ( B ) Quantification analysis of Topo I activity. ( C ) Topo I protein level examined by Western blot analysis. Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture t -test: * p

    Techniques Used: Cycling Probe Technology, Inhibition, Activity Assay, Infection, Agarose Gel Electrophoresis, Western Blot

    The Effect of MEK inhibitor on Mycoplasma-induced inhibitory effect on Topo I activity and on ERK 1/2 Phosphorylation in MCF7 cells. MCF7 cells were pre-incubated with MEK inhibitors (PD) for 1 hour at a concentration of 25 µM followed by M. fermentans infection (MOI of 10 3 CFU/cell) for an additional 6 hrs. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis (A) and Topo I activity was quantified (B). Phosphorylated ERK 1/2 protein level from MCF7-extract was examined by Western blot analysis (C). Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture t -test: *** p
    Figure Legend Snippet: The Effect of MEK inhibitor on Mycoplasma-induced inhibitory effect on Topo I activity and on ERK 1/2 Phosphorylation in MCF7 cells. MCF7 cells were pre-incubated with MEK inhibitors (PD) for 1 hour at a concentration of 25 µM followed by M. fermentans infection (MOI of 10 3 CFU/cell) for an additional 6 hrs. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis (A) and Topo I activity was quantified (B). Phosphorylated ERK 1/2 protein level from MCF7-extract was examined by Western blot analysis (C). Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture t -test: *** p

    Techniques Used: Activity Assay, Incubation, Concentration Assay, Infection, Agarose Gel Electrophoresis, Western Blot

    Related Articles

    Agarose Gel Electrophoresis:

    Article Title: Glutamate Activates AMPA Receptor Conductance in the Developing Schwann Cells of the Mammalian Peripheral Nerves
    Article Snippet: .. The digestion products were analyzed by agarose gel electrophoresis (stained with GelRel) with the marker pUC19 DNA/MspI (Hpall) (Thermo Scientific). .. For positive controls, HEK293 were transfected with pRK5-GluR1i-L497Y-(Ser831-Ser845), pRetroCAG-GluA2(Q583R), pRK5-GluA3 flip, or pCI-EGFPGluA4 using lipofectamine 2000 (Invitrogen) according to the manufacturer's protocol.

    Concentration Assay:

    Article Title: DNA Damage-Response Pathway Heterogeneity of Human Lung Cancer A549 and H1299 Cells Determines Sensitivity to 8-Chloro-Adenosine
    Article Snippet: .. DNA Relaxation Reaction mixtures containing 0.4 mg pUC19 plasmid DNA (MBI Fermentas, Vilnius, Lithuania) and 2.5 μg nuclear extracts (NE) from 8-Cl-Ado-exposed or -unexposed cells, or 5 mM 3-aminobenzamide (PARP inhibitor) in 20 μL relaxation buffer (50 mM Tris-HCl, pH 8.0, 0.1 M NaCl, 5 mM MgCl2 ) were incubated at 37 °C for 30 min and stopped by adding sodium dodecyl sulphate (SDS) and ethylenediaminetetraacetic acid (EDTA) to a final concentration of 0.1% and 10 mM, respectively. .. DNA was ethanol precipitated, and subjected to electrophoresis in 1% agarose gels.

    Incubation:

    Article Title: DNA Damage-Response Pathway Heterogeneity of Human Lung Cancer A549 and H1299 Cells Determines Sensitivity to 8-Chloro-Adenosine
    Article Snippet: .. DNA Relaxation Reaction mixtures containing 0.4 mg pUC19 plasmid DNA (MBI Fermentas, Vilnius, Lithuania) and 2.5 μg nuclear extracts (NE) from 8-Cl-Ado-exposed or -unexposed cells, or 5 mM 3-aminobenzamide (PARP inhibitor) in 20 μL relaxation buffer (50 mM Tris-HCl, pH 8.0, 0.1 M NaCl, 5 mM MgCl2 ) were incubated at 37 °C for 30 min and stopped by adding sodium dodecyl sulphate (SDS) and ethylenediaminetetraacetic acid (EDTA) to a final concentration of 0.1% and 10 mM, respectively. .. DNA was ethanol precipitated, and subjected to electrophoresis in 1% agarose gels.

    Marker:

    Article Title: Glutamate Activates AMPA Receptor Conductance in the Developing Schwann Cells of the Mammalian Peripheral Nerves
    Article Snippet: .. The digestion products were analyzed by agarose gel electrophoresis (stained with GelRel) with the marker pUC19 DNA/MspI (Hpall) (Thermo Scientific). .. For positive controls, HEK293 were transfected with pRK5-GluR1i-L497Y-(Ser831-Ser845), pRetroCAG-GluA2(Q583R), pRK5-GluA3 flip, or pCI-EGFPGluA4 using lipofectamine 2000 (Invitrogen) according to the manufacturer's protocol.

    Staining:

    Article Title: Glutamate Activates AMPA Receptor Conductance in the Developing Schwann Cells of the Mammalian Peripheral Nerves
    Article Snippet: .. The digestion products were analyzed by agarose gel electrophoresis (stained with GelRel) with the marker pUC19 DNA/MspI (Hpall) (Thermo Scientific). .. For positive controls, HEK293 were transfected with pRK5-GluR1i-L497Y-(Ser831-Ser845), pRetroCAG-GluA2(Q583R), pRK5-GluA3 flip, or pCI-EGFPGluA4 using lipofectamine 2000 (Invitrogen) according to the manufacturer's protocol.

    Plasmid Preparation:

    Article Title: The structural characterization of a prophage-encoded extracellular DNase from Streptococcus pyogenes
    Article Snippet: .. Preparation of DNA substrate The plasmid vector pUC19 was linearized with EcoRI (Fermentas) and used as a homogeneous substrate for the continuous assay. .. The linearization was verified by agarose gel electrophoresis, and the restriction endonuclease inactivated in accordance with the manufacturer's instructions (heat denaturation at 65°C for 20 min).

    Article Title: An artificial metalloenzyme for catalytic cancer-specific DNA cleavage and operando imaging
    Article Snippet: .. The Supercoiled pUC19 plasmid DNA with 2686 bp was purchased from Thermo Fisher Scientific for the DNA double-strand break assays. .. Cell culture 1640 medium and fetal bovine serum (FBS) were purchased from HyClone.

    Article Title: DNA Damage-Response Pathway Heterogeneity of Human Lung Cancer A549 and H1299 Cells Determines Sensitivity to 8-Chloro-Adenosine
    Article Snippet: .. DNA Relaxation Reaction mixtures containing 0.4 mg pUC19 plasmid DNA (MBI Fermentas, Vilnius, Lithuania) and 2.5 μg nuclear extracts (NE) from 8-Cl-Ado-exposed or -unexposed cells, or 5 mM 3-aminobenzamide (PARP inhibitor) in 20 μL relaxation buffer (50 mM Tris-HCl, pH 8.0, 0.1 M NaCl, 5 mM MgCl2 ) were incubated at 37 °C for 30 min and stopped by adding sodium dodecyl sulphate (SDS) and ethylenediaminetetraacetic acid (EDTA) to a final concentration of 0.1% and 10 mM, respectively. .. DNA was ethanol precipitated, and subjected to electrophoresis in 1% agarose gels.

    Article Title: Evaluation of digital real-time PCR assay as a molecular diagnostic tool for single-cell analysis
    Article Snippet: .. The pUC19 plasmid DNA standard was quantitated using Nanodrop™ (Thermo Fisher Scientific, Taiwan) and converted into the DNA copy number used in the calculations in this study. ..

    Article Title: Mycoplasma fermentans Inhibits the Activity of Cellular DNA Topoisomerase I by Activation of PARP1 and Alters the Efficacy of Its Anti-Cancer Inhibitor
    Article Snippet: .. Supercoiled DNA plasmid pUC19 was purchased from MBI Fermentas (Hanover, MD, USA). .. PD98059 and 3-aminobenzamide (3AB) were purchased from Sigma-Aldrich (Rehovot, Israel).

    Article Title: The Post-Synaptic Function of Brca2
    Article Snippet: .. DNA Unwinding Assay Relaxed DNA was prepared in a 50 µl reaction mixture by incubating negatively supercoiled plasmid pUC19 DNA (0.5 µg) with calf thymus Topoisomerase I (1 Unit; Invitrogen) in buffer containing 50 mM Tris HCl pH 7.5, 50 mM KCl, 10 mM MgCl2 , 0.5 mM DTT, 0.1 mM EDTA, 100 µg ml−1 BSA for 30 min at 37 °C. .. To test the unwinding activity of RAD51, RAD51 K133A, RAD51 K133R, these proteins were incubated with relaxed pUC19 (25 µM, nt) and unlabeled #160 (12.5 µM, nt) in the D-loop formation buffer supplemented with 10 mM magnesium acetate for 5 min at 37 °C.

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  • 98
    Thermo Fisher supercoiled puc19 plasmid dna
    Copper clusters as anticancer biomimetic catalysts exert therapeutic effect in vitro and in vivo. ( A ) In vitro viability of A549 lung tumor cells after incubation of RGD-BSA-CuCs or BSA-CuCs with the identical clusters concentration for 48 hours. Viability of A549 cells pretreated by RGD-BSA is set as control group (means ± SD, n = 3). DIC, differential interference contrast. ( B ) Fluorescence microscopy images showing ROS burst through fluorogenic reaction with CM-H 2 DCFDA in cells pretreated with a series concentration of RGD-BSA-CuCs for 48 hours. Cells treated with 39 μM RGD-BSA-CuCs and 250 μM ROS quencher mannitol are set as control. ( C ) Quantification the proportion of apoptotic cells in series dosages of RGD-BSA-CuCs measured by flow cytometry. DIC, differential interference contrast. ( D ) Agarose gel electrophoretic patterns of <t>supercoiled</t> pUC19 plasmid DNA (20 ng ml −1 ) in the presence of a series concentration of RGD-BSA-CuCs and 100 μM H 2 O 2 (left). The mixture was preincubated in pH 7.4 buffer solution at 37°C for 12 hours. The persistent DNA cleavage property of 9.6 μM RGD-BSA-CuCs in the presence of 100 μM H 2 O 2 (right). The mixture was preincubated in pH 7.4 buffer solution at 37°C for 0, 3, 6, and 12 hours. DNA treated by 100 μM H 2 O 2 for 12 hours is set as control. ( E ) DNA lesions induced by the increased concentration of RGD-BSA-CuCs measured by the basic comet assay. DNA content in the comet tail represents the extent of DNA cleavage, analyzed by 50 cells selected randomly by CASP software. *** P
    Supercoiled Puc19 Plasmid Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 0 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/supercoiled puc19 plasmid dna/product/Thermo Fisher
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    supercoiled puc19 plasmid dna - by Bioz Stars, 2020-09
    98/100 stars
      Buy from Supplier

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    Copper clusters as anticancer biomimetic catalysts exert therapeutic effect in vitro and in vivo. ( A ) In vitro viability of A549 lung tumor cells after incubation of RGD-BSA-CuCs or BSA-CuCs with the identical clusters concentration for 48 hours. Viability of A549 cells pretreated by RGD-BSA is set as control group (means ± SD, n = 3). DIC, differential interference contrast. ( B ) Fluorescence microscopy images showing ROS burst through fluorogenic reaction with CM-H 2 DCFDA in cells pretreated with a series concentration of RGD-BSA-CuCs for 48 hours. Cells treated with 39 μM RGD-BSA-CuCs and 250 μM ROS quencher mannitol are set as control. ( C ) Quantification the proportion of apoptotic cells in series dosages of RGD-BSA-CuCs measured by flow cytometry. DIC, differential interference contrast. ( D ) Agarose gel electrophoretic patterns of supercoiled pUC19 plasmid DNA (20 ng ml −1 ) in the presence of a series concentration of RGD-BSA-CuCs and 100 μM H 2 O 2 (left). The mixture was preincubated in pH 7.4 buffer solution at 37°C for 12 hours. The persistent DNA cleavage property of 9.6 μM RGD-BSA-CuCs in the presence of 100 μM H 2 O 2 (right). The mixture was preincubated in pH 7.4 buffer solution at 37°C for 0, 3, 6, and 12 hours. DNA treated by 100 μM H 2 O 2 for 12 hours is set as control. ( E ) DNA lesions induced by the increased concentration of RGD-BSA-CuCs measured by the basic comet assay. DNA content in the comet tail represents the extent of DNA cleavage, analyzed by 50 cells selected randomly by CASP software. *** P

    Journal: Science Advances

    Article Title: An artificial metalloenzyme for catalytic cancer-specific DNA cleavage and operando imaging

    doi: 10.1126/sciadv.abb1421

    Figure Lengend Snippet: Copper clusters as anticancer biomimetic catalysts exert therapeutic effect in vitro and in vivo. ( A ) In vitro viability of A549 lung tumor cells after incubation of RGD-BSA-CuCs or BSA-CuCs with the identical clusters concentration for 48 hours. Viability of A549 cells pretreated by RGD-BSA is set as control group (means ± SD, n = 3). DIC, differential interference contrast. ( B ) Fluorescence microscopy images showing ROS burst through fluorogenic reaction with CM-H 2 DCFDA in cells pretreated with a series concentration of RGD-BSA-CuCs for 48 hours. Cells treated with 39 μM RGD-BSA-CuCs and 250 μM ROS quencher mannitol are set as control. ( C ) Quantification the proportion of apoptotic cells in series dosages of RGD-BSA-CuCs measured by flow cytometry. DIC, differential interference contrast. ( D ) Agarose gel electrophoretic patterns of supercoiled pUC19 plasmid DNA (20 ng ml −1 ) in the presence of a series concentration of RGD-BSA-CuCs and 100 μM H 2 O 2 (left). The mixture was preincubated in pH 7.4 buffer solution at 37°C for 12 hours. The persistent DNA cleavage property of 9.6 μM RGD-BSA-CuCs in the presence of 100 μM H 2 O 2 (right). The mixture was preincubated in pH 7.4 buffer solution at 37°C for 0, 3, 6, and 12 hours. DNA treated by 100 μM H 2 O 2 for 12 hours is set as control. ( E ) DNA lesions induced by the increased concentration of RGD-BSA-CuCs measured by the basic comet assay. DNA content in the comet tail represents the extent of DNA cleavage, analyzed by 50 cells selected randomly by CASP software. *** P

    Article Snippet: The Supercoiled pUC19 plasmid DNA with 2686 bp was purchased from Thermo Fisher Scientific for the DNA double-strand break assays.

    Techniques: In Vitro, In Vivo, Incubation, Concentration Assay, Fluorescence, Microscopy, Flow Cytometry, Agarose Gel Electrophoresis, Plasmid Preparation, Single Cell Gel Electrophoresis, Software