hepes  (Gold Biotechnology Inc)


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    Structured Review

    Gold Biotechnology Inc hepes
    (A) Cyclic voltammograms of WT Mn/Fe R2lox from pH 6 to pH 9 (as indicated). (Inset) Anodic (open symbols) and cathodic (closed symbols) peak potentials for WT Mn/Fe R2lox as a function of pH. Dotted lines indicate a slope of −120 mV/pH unit. (B) Cyclic voltammograms of Mn/Fe (solid) and <t>apo</t> (dashed) R2lox at pH 7.0. (C) Anodic (open symbols) and cathodic (closed symbols) peak potentials as a function of scan rate. All samples contained 100 μ M WT (black), Y175F (blue), or E69D (green) R2lox protein with 100 mM NaCl in 100 mM <t>HEPES</t> buffer, pH 8.0. Points and error bars reflect the average and standard deviation of n ≥ 3 trials, respectively. Samples measured with a glassy carbon working electrode.
    Hepes, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 94/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Gold Biotechnology Inc
    Average 94 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2022-08
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    Images

    1) Product Images from "Key Structural Motifs Balance Metal Binding and Oxidative Reactivity in a Heterobimetallic Mn/Fe Protein"

    Article Title: Key Structural Motifs Balance Metal Binding and Oxidative Reactivity in a Heterobimetallic Mn/Fe Protein

    Journal: Journal of the American Chemical Society

    doi: 10.1021/jacs.0c00333

    (A) Cyclic voltammograms of WT Mn/Fe R2lox from pH 6 to pH 9 (as indicated). (Inset) Anodic (open symbols) and cathodic (closed symbols) peak potentials for WT Mn/Fe R2lox as a function of pH. Dotted lines indicate a slope of −120 mV/pH unit. (B) Cyclic voltammograms of Mn/Fe (solid) and apo (dashed) R2lox at pH 7.0. (C) Anodic (open symbols) and cathodic (closed symbols) peak potentials as a function of scan rate. All samples contained 100 μ M WT (black), Y175F (blue), or E69D (green) R2lox protein with 100 mM NaCl in 100 mM HEPES buffer, pH 8.0. Points and error bars reflect the average and standard deviation of n ≥ 3 trials, respectively. Samples measured with a glassy carbon working electrode.
    Figure Legend Snippet: (A) Cyclic voltammograms of WT Mn/Fe R2lox from pH 6 to pH 9 (as indicated). (Inset) Anodic (open symbols) and cathodic (closed symbols) peak potentials for WT Mn/Fe R2lox as a function of pH. Dotted lines indicate a slope of −120 mV/pH unit. (B) Cyclic voltammograms of Mn/Fe (solid) and apo (dashed) R2lox at pH 7.0. (C) Anodic (open symbols) and cathodic (closed symbols) peak potentials as a function of scan rate. All samples contained 100 μ M WT (black), Y175F (blue), or E69D (green) R2lox protein with 100 mM NaCl in 100 mM HEPES buffer, pH 8.0. Points and error bars reflect the average and standard deviation of n ≥ 3 trials, respectively. Samples measured with a glassy carbon working electrode.

    Techniques Used: Standard Deviation

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    Gold Biotechnology Inc hygromycin b
    Schematic of CRISPR-Cas9 mediated gene deletion, cassette integration, and excision. (A) GENECC9KO primer pairs are designed to bind to conserved regions on SAT1 - and CaHygB -flipper plasmids (black lines) and also contain gene-specific overhangs (red lines) that are complementary to the target gene of interest and located close to PAM sites in the C. albicans genome that flank the gene targeted for deletion. GENECC9KO primers are used to generate repair templates simultaneously utilized during CRISPR-Cas9-mediated transformation. (B) After transformation and selection on media containing both NAT and <t>HygB,</t> each cassette integrates at each target gene locus to generate homozygous deletion mutants. (C) After growth in maltose medium (YPM), the cassettes are “flipped” out by flip recombinase mediated cleavage at FRT sites. A single FRT site and approximately 200 bp of additional plasmid sequence (termed FRT+) remain at each locus. Primer pairs FLIPINTF and GENEINTR and FLPINTF and FLPINTR are used to detect locus disruption by PCR. Loss of the coding sequence is also confirmed by GENEDET primers.
    Hygromycin B, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Gold Biotechnology Inc hepes
    (A) Cyclic voltammograms of WT Mn/Fe R2lox from pH 6 to pH 9 (as indicated). (Inset) Anodic (open symbols) and cathodic (closed symbols) peak potentials for WT Mn/Fe R2lox as a function of pH. Dotted lines indicate a slope of −120 mV/pH unit. (B) Cyclic voltammograms of Mn/Fe (solid) and <t>apo</t> (dashed) R2lox at pH 7.0. (C) Anodic (open symbols) and cathodic (closed symbols) peak potentials as a function of scan rate. All samples contained 100 μ M WT (black), Y175F (blue), or E69D (green) R2lox protein with 100 mM NaCl in 100 mM <t>HEPES</t> buffer, pH 8.0. Points and error bars reflect the average and standard deviation of n ≥ 3 trials, respectively. Samples measured with a glassy carbon working electrode.
    Hepes, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Gold Biotechnology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2022-08
    94/100 stars
      Buy from Supplier

    Image Search Results


    Schematic of CRISPR-Cas9 mediated gene deletion, cassette integration, and excision. (A) GENECC9KO primer pairs are designed to bind to conserved regions on SAT1 - and CaHygB -flipper plasmids (black lines) and also contain gene-specific overhangs (red lines) that are complementary to the target gene of interest and located close to PAM sites in the C. albicans genome that flank the gene targeted for deletion. GENECC9KO primers are used to generate repair templates simultaneously utilized during CRISPR-Cas9-mediated transformation. (B) After transformation and selection on media containing both NAT and HygB, each cassette integrates at each target gene locus to generate homozygous deletion mutants. (C) After growth in maltose medium (YPM), the cassettes are “flipped” out by flip recombinase mediated cleavage at FRT sites. A single FRT site and approximately 200 bp of additional plasmid sequence (termed FRT+) remain at each locus. Primer pairs FLIPINTF and GENEINTR and FLPINTF and FLPINTR are used to detect locus disruption by PCR. Loss of the coding sequence is also confirmed by GENEDET primers.

    Journal: Microbiology Spectrum

    Article Title: Rapid Hypothesis Testing in Candida albicans Clinical Isolates Using a Cloning-Free, Modular, and Recyclable System for CRISPR-Cas9 Mediated Mutant and Revertant Construction

    doi: 10.1128/spectrum.02630-21

    Figure Lengend Snippet: Schematic of CRISPR-Cas9 mediated gene deletion, cassette integration, and excision. (A) GENECC9KO primer pairs are designed to bind to conserved regions on SAT1 - and CaHygB -flipper plasmids (black lines) and also contain gene-specific overhangs (red lines) that are complementary to the target gene of interest and located close to PAM sites in the C. albicans genome that flank the gene targeted for deletion. GENECC9KO primers are used to generate repair templates simultaneously utilized during CRISPR-Cas9-mediated transformation. (B) After transformation and selection on media containing both NAT and HygB, each cassette integrates at each target gene locus to generate homozygous deletion mutants. (C) After growth in maltose medium (YPM), the cassettes are “flipped” out by flip recombinase mediated cleavage at FRT sites. A single FRT site and approximately 200 bp of additional plasmid sequence (termed FRT+) remain at each locus. Primer pairs FLIPINTF and GENEINTR and FLPINTF and FLPINTR are used to detect locus disruption by PCR. Loss of the coding sequence is also confirmed by GENEDET primers.

    Article Snippet: Several semilog dilutions were prepared in TE buffer prior to spread-plating 100 μL of each suspension on YPD plates containing a range of nourseothricin (75 – 200 μg/mL, GoldBio) and hygromycin B (400–700 μg/mL, GoldBio).

    Techniques: CRISPR, Transformation Assay, Selection, Plasmid Preparation, Sequencing, Polymerase Chain Reaction

    Homozygous gene targeting of the ADE2 locus using CRISPR-Cas9 is highly efficient across diverse clinical isolates. (A) Transformation efficiency was calculated for strains SC5314 (bloodstream isolate and reference strain), 529L (oral isolate), JS15 (vaginal isolate), SJCA1 (catheter isolate), and TW1 (oral isolate) and plotted as mean ± SD of 3 independent experiments. Data were assessed for statistical significance by one-way ANOVA and Tukey’s posttest. ns, not significant (B) Representative images of YPD plates containing 200 μg/mL NAT and 600 μg/mL HygB were digitally captured. (C) Marker excision for strains JS15, SJCA1, 529L, and TW1 (not shown) were validated by patching resulting colonies after overnight growth in YPM medium and selection onto medium containing 200 μg/mL NAT, 600 μg/mL, or no antimicrobial.

    Journal: Microbiology Spectrum

    Article Title: Rapid Hypothesis Testing in Candida albicans Clinical Isolates Using a Cloning-Free, Modular, and Recyclable System for CRISPR-Cas9 Mediated Mutant and Revertant Construction

    doi: 10.1128/spectrum.02630-21

    Figure Lengend Snippet: Homozygous gene targeting of the ADE2 locus using CRISPR-Cas9 is highly efficient across diverse clinical isolates. (A) Transformation efficiency was calculated for strains SC5314 (bloodstream isolate and reference strain), 529L (oral isolate), JS15 (vaginal isolate), SJCA1 (catheter isolate), and TW1 (oral isolate) and plotted as mean ± SD of 3 independent experiments. Data were assessed for statistical significance by one-way ANOVA and Tukey’s posttest. ns, not significant (B) Representative images of YPD plates containing 200 μg/mL NAT and 600 μg/mL HygB were digitally captured. (C) Marker excision for strains JS15, SJCA1, 529L, and TW1 (not shown) were validated by patching resulting colonies after overnight growth in YPM medium and selection onto medium containing 200 μg/mL NAT, 600 μg/mL, or no antimicrobial.

    Article Snippet: Several semilog dilutions were prepared in TE buffer prior to spread-plating 100 μL of each suspension on YPD plates containing a range of nourseothricin (75 – 200 μg/mL, GoldBio) and hygromycin B (400–700 μg/mL, GoldBio).

    Techniques: CRISPR, Transformation Assay, Marker, Selection

    Schematic of CaHygB -flipper plasmid construction. (A) A segment containing the flip recombinase ( FLP ), ACT1 terminator, and ACT1 promoter of the SAT1 -flipper plasmid was PCR amplified and (B) ligated into SalI and PstI digested SAT1 -flipper. This resulted in plasmid pBSS2-Δ SAT1 which lacked the SAT1 gene encoding resistance to nourseothricin. (C) A synthetic piece of DNA encoding a C. albicans codon optimized hygromycin B resistance gene ( CaHygB ) and URA3 terminator was ligated into pBSS2-Δ SAT1 to create the CaHygB -flipper plasmid.

    Journal: Microbiology Spectrum

    Article Title: Rapid Hypothesis Testing in Candida albicans Clinical Isolates Using a Cloning-Free, Modular, and Recyclable System for CRISPR-Cas9 Mediated Mutant and Revertant Construction

    doi: 10.1128/spectrum.02630-21

    Figure Lengend Snippet: Schematic of CaHygB -flipper plasmid construction. (A) A segment containing the flip recombinase ( FLP ), ACT1 terminator, and ACT1 promoter of the SAT1 -flipper plasmid was PCR amplified and (B) ligated into SalI and PstI digested SAT1 -flipper. This resulted in plasmid pBSS2-Δ SAT1 which lacked the SAT1 gene encoding resistance to nourseothricin. (C) A synthetic piece of DNA encoding a C. albicans codon optimized hygromycin B resistance gene ( CaHygB ) and URA3 terminator was ligated into pBSS2-Δ SAT1 to create the CaHygB -flipper plasmid.

    Article Snippet: Several semilog dilutions were prepared in TE buffer prior to spread-plating 100 μL of each suspension on YPD plates containing a range of nourseothricin (75 – 200 μg/mL, GoldBio) and hygromycin B (400–700 μg/mL, GoldBio).

    Techniques: Plasmid Preparation, Polymerase Chain Reaction, Amplification

    (A) Cyclic voltammograms of WT Mn/Fe R2lox from pH 6 to pH 9 (as indicated). (Inset) Anodic (open symbols) and cathodic (closed symbols) peak potentials for WT Mn/Fe R2lox as a function of pH. Dotted lines indicate a slope of −120 mV/pH unit. (B) Cyclic voltammograms of Mn/Fe (solid) and apo (dashed) R2lox at pH 7.0. (C) Anodic (open symbols) and cathodic (closed symbols) peak potentials as a function of scan rate. All samples contained 100 μ M WT (black), Y175F (blue), or E69D (green) R2lox protein with 100 mM NaCl in 100 mM HEPES buffer, pH 8.0. Points and error bars reflect the average and standard deviation of n ≥ 3 trials, respectively. Samples measured with a glassy carbon working electrode.

    Journal: Journal of the American Chemical Society

    Article Title: Key Structural Motifs Balance Metal Binding and Oxidative Reactivity in a Heterobimetallic Mn/Fe Protein

    doi: 10.1021/jacs.0c00333

    Figure Lengend Snippet: (A) Cyclic voltammograms of WT Mn/Fe R2lox from pH 6 to pH 9 (as indicated). (Inset) Anodic (open symbols) and cathodic (closed symbols) peak potentials for WT Mn/Fe R2lox as a function of pH. Dotted lines indicate a slope of −120 mV/pH unit. (B) Cyclic voltammograms of Mn/Fe (solid) and apo (dashed) R2lox at pH 7.0. (C) Anodic (open symbols) and cathodic (closed symbols) peak potentials as a function of scan rate. All samples contained 100 μ M WT (black), Y175F (blue), or E69D (green) R2lox protein with 100 mM NaCl in 100 mM HEPES buffer, pH 8.0. Points and error bars reflect the average and standard deviation of n ≥ 3 trials, respectively. Samples measured with a glassy carbon working electrode.

    Article Snippet: Apo-R2lox aliquots were thawed and diluted into 100 mM HEPES (Goldbio) buffer, pH 7.0, containing 50 mM NaCl (Fisher Scientific), subsequently referred to as “Buffer E”.

    Techniques: Standard Deviation