Structured Review

Millipore tris cl
Release of glyoxal from the dihydro-dihydroxy form derived from <t>benznidazole.</t> (A) Schematic showing the release of glyoxal from dihydro-dihydroxyimidazole. (B) Spectrophotometric detection of glyoxal in reaction mixtures containing TbNTR (40 μg), benznidazole (200 μM), and various quantities of NADH (0 to 800 μM) in 1 ml 50 mM NaH 2 PO 4 (pH 7.5). To detect glyoxal, an aliquot of the reaction mixture was added to a borate buffer (pH 9.2) containing Girard T reagent, and the absorbance of the mixture at 325 nm was then determined. The concentration of glyoxal in the reaction mixtures was calculated by comparison against a standard curve obtained by using pure dialdehyde, after the subtraction of blank readings from samples lacking Girard T reagent. Values are means derived from experiments performed in triplicate ± standard deviations. (C) Time-dependent production of glyoxal from benznidazole. A 1-ml reaction mixture containing benznidazole (200 μM) in 50 mM <t>Tris-Cl</t> (pH 7.5) was incubated at room temperature with no addition (●), NADH (total amount added, 800 μM) (■), or NADH (total amount added, 800 μM) and 4 μg TbNTR (▴). At each time point, aliquots (100 μl) were taken and assayed for the presence of glyoxal. Glyoxal concentrations were then calculated as described above.
Tris Cl, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/tris cl/product/Millipore
Average 97 stars, based on 1 article reviews
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tris cl - by Bioz Stars, 2022-01
97/100 stars

Images

1) Product Images from "Activation of Benznidazole by Trypanosomal Type I Nitroreductases Results in Glyoxal Formation"

Article Title: Activation of Benznidazole by Trypanosomal Type I Nitroreductases Results in Glyoxal Formation

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.05135-11

Release of glyoxal from the dihydro-dihydroxy form derived from benznidazole. (A) Schematic showing the release of glyoxal from dihydro-dihydroxyimidazole. (B) Spectrophotometric detection of glyoxal in reaction mixtures containing TbNTR (40 μg), benznidazole (200 μM), and various quantities of NADH (0 to 800 μM) in 1 ml 50 mM NaH 2 PO 4 (pH 7.5). To detect glyoxal, an aliquot of the reaction mixture was added to a borate buffer (pH 9.2) containing Girard T reagent, and the absorbance of the mixture at 325 nm was then determined. The concentration of glyoxal in the reaction mixtures was calculated by comparison against a standard curve obtained by using pure dialdehyde, after the subtraction of blank readings from samples lacking Girard T reagent. Values are means derived from experiments performed in triplicate ± standard deviations. (C) Time-dependent production of glyoxal from benznidazole. A 1-ml reaction mixture containing benznidazole (200 μM) in 50 mM Tris-Cl (pH 7.5) was incubated at room temperature with no addition (●), NADH (total amount added, 800 μM) (■), or NADH (total amount added, 800 μM) and 4 μg TbNTR (▴). At each time point, aliquots (100 μl) were taken and assayed for the presence of glyoxal. Glyoxal concentrations were then calculated as described above.
Figure Legend Snippet: Release of glyoxal from the dihydro-dihydroxy form derived from benznidazole. (A) Schematic showing the release of glyoxal from dihydro-dihydroxyimidazole. (B) Spectrophotometric detection of glyoxal in reaction mixtures containing TbNTR (40 μg), benznidazole (200 μM), and various quantities of NADH (0 to 800 μM) in 1 ml 50 mM NaH 2 PO 4 (pH 7.5). To detect glyoxal, an aliquot of the reaction mixture was added to a borate buffer (pH 9.2) containing Girard T reagent, and the absorbance of the mixture at 325 nm was then determined. The concentration of glyoxal in the reaction mixtures was calculated by comparison against a standard curve obtained by using pure dialdehyde, after the subtraction of blank readings from samples lacking Girard T reagent. Values are means derived from experiments performed in triplicate ± standard deviations. (C) Time-dependent production of glyoxal from benznidazole. A 1-ml reaction mixture containing benznidazole (200 μM) in 50 mM Tris-Cl (pH 7.5) was incubated at room temperature with no addition (●), NADH (total amount added, 800 μM) (■), or NADH (total amount added, 800 μM) and 4 μg TbNTR (▴). At each time point, aliquots (100 μl) were taken and assayed for the presence of glyoxal. Glyoxal concentrations were then calculated as described above.

Techniques Used: Derivative Assay, Concentration Assay, Incubation

2) Product Images from "Functional role of PGAM5 multimeric assemblies and their polymerization into filaments"

Article Title: Functional role of PGAM5 multimeric assemblies and their polymerization into filaments

Journal: Nature Communications

doi: 10.1038/s41467-019-08393-w

∆48 PGAM5 forms tubular filaments in solution composed of ring-like structures. a Domain architecture of constructs used in this study. Full-length PGAM5 is comprised of a single transmembrane helix containing a mitochondrial targeting sequence (MTS), a linker domain including the regulatory multimerization motif (MM), and a C-terminal PGAM phosphatase domain. The native cleavage site between residues 24 and 25, cleaved by PARL 6 , is marked with a green arrow. b Elution profiles for ∆48 and ∆90 PGAM5 constructs purified by size exclusion chromatography (SEC) using a Superose 6 column (GE Healthcare) in SEC buffer containing 20 mM Tris-Cl pH 8.0, 500 µM TCEP, and 150 mM NaCl. The corresponding oligomeric states of each peak observed in the chromatograms are indicated. c Representative EM micrographs of negatively stained protein samples taken from fractions corresponding to the two distinct peaks observed in the ∆48 PGAM5 purification. d SEC profiles for ∆48 PGAM5 in SEC buffer and NaCl at a final concentration of 150, 300, or 750 mM. EM micrographs of negatively stained samples of ∆48 PGAM5 taken directly from the primary peak obtained during purification are shown, highlighting the decomposition of filaments into rings at increasing salt concentrations. Scale bars in c and d correspond to 50 nm, except for the inset in c in which the scale bar corresponds to 10 nm
Figure Legend Snippet: ∆48 PGAM5 forms tubular filaments in solution composed of ring-like structures. a Domain architecture of constructs used in this study. Full-length PGAM5 is comprised of a single transmembrane helix containing a mitochondrial targeting sequence (MTS), a linker domain including the regulatory multimerization motif (MM), and a C-terminal PGAM phosphatase domain. The native cleavage site between residues 24 and 25, cleaved by PARL 6 , is marked with a green arrow. b Elution profiles for ∆48 and ∆90 PGAM5 constructs purified by size exclusion chromatography (SEC) using a Superose 6 column (GE Healthcare) in SEC buffer containing 20 mM Tris-Cl pH 8.0, 500 µM TCEP, and 150 mM NaCl. The corresponding oligomeric states of each peak observed in the chromatograms are indicated. c Representative EM micrographs of negatively stained protein samples taken from fractions corresponding to the two distinct peaks observed in the ∆48 PGAM5 purification. d SEC profiles for ∆48 PGAM5 in SEC buffer and NaCl at a final concentration of 150, 300, or 750 mM. EM micrographs of negatively stained samples of ∆48 PGAM5 taken directly from the primary peak obtained during purification are shown, highlighting the decomposition of filaments into rings at increasing salt concentrations. Scale bars in c and d correspond to 50 nm, except for the inset in c in which the scale bar corresponds to 10 nm

Techniques Used: Construct, Sequencing, Purification, Size-exclusion Chromatography, Staining, Concentration Assay

3) Product Images from "Nucleosome Recognition by the Piccolo NuA4 Histone Acetyltransferase Complex †"

Article Title: Nucleosome Recognition by the Piccolo NuA4 Histone Acetyltransferase Complex †

Journal: Biochemistry

doi: 10.1021/bi602366n

Substrate specificity of picNuA4. All reactions contained 1 × TBA at pH 7.0 or 50 mM Tris at pH 7.2, 1 mM DTT, 50–100 μ M acetyl-CoA, and 0.01–0.1 μ M picNuA4 complex. Concentrations of peptide substrates used in saturation curves were varied over a 10-fold range covering concentrations above and below the K m . Data were fitted to appropriate equations in Kaleidagraph to determine the kinetic constants. Each value is an average of three to six separate assays, with the standard deviation shown. Gray bars represent values from saturation curves ( k cat / K m ), and black bars represent values determined from progress curves ( k cat / K avg ). (***) Value represents the lower limit for initial k cat / K m of NCPs because data below 150 nM were not collected.
Figure Legend Snippet: Substrate specificity of picNuA4. All reactions contained 1 × TBA at pH 7.0 or 50 mM Tris at pH 7.2, 1 mM DTT, 50–100 μ M acetyl-CoA, and 0.01–0.1 μ M picNuA4 complex. Concentrations of peptide substrates used in saturation curves were varied over a 10-fold range covering concentrations above and below the K m . Data were fitted to appropriate equations in Kaleidagraph to determine the kinetic constants. Each value is an average of three to six separate assays, with the standard deviation shown. Gray bars represent values from saturation curves ( k cat / K m ), and black bars represent values determined from progress curves ( k cat / K avg ). (***) Value represents the lower limit for initial k cat / K m of NCPs because data below 150 nM were not collected.

Techniques Used: Standard Deviation

Substrate specificity of Esa1. All reactions contain 1 × TBA at pH 7.0 or 50 mM Tris at pH 7.2, 1 mM DTT, 75–100 μ M acetyl-CoA, and 0.54–2 μ M Esa1. Concentrations of peptide substrates in saturation curves were varied over a 10-fold range covering concentrations above and below the K m . Data were fitted to the Michaelis–Menten equation in Kaleidagraph to determine kinetic constants. The k cat / K m value for nucleosome arrays was collected under subsaturating conditions, and accordingly, the data were fitted to a linear equation where the slope is k cat / K m.
Figure Legend Snippet: Substrate specificity of Esa1. All reactions contain 1 × TBA at pH 7.0 or 50 mM Tris at pH 7.2, 1 mM DTT, 75–100 μ M acetyl-CoA, and 0.54–2 μ M Esa1. Concentrations of peptide substrates in saturation curves were varied over a 10-fold range covering concentrations above and below the K m . Data were fitted to the Michaelis–Menten equation in Kaleidagraph to determine kinetic constants. The k cat / K m value for nucleosome arrays was collected under subsaturating conditions, and accordingly, the data were fitted to a linear equation where the slope is k cat / K m.

Techniques Used:

4) Product Images from "A novel FOXA1/ESR1 interacting pathway: A study of Oncomine™ breast cancer microarrays"

Article Title: A novel FOXA1/ESR1 interacting pathway: A study of Oncomine™ breast cancer microarrays

Journal: Oncology Letters

doi: 10.3892/ol.2017.6329

Schematic representation of the PS2 promoter. (A) Schematic diagram showing the presence of a functional estrogen response element (−407 nucleotide position) and two putative FOXA1 binding sites at −384 and −539 nucleotide positions, respectively. (B) In vitro binding assay. A total of 30 bp oligonucleotides containing FOXA1 binding sites were labeled with γ 32 P radioisotope and incubated with nuclear lysate extracted from MCF7 cells. An unlabeled FOXA1 (cold probe) consensus sequence was used for competition at 100 and 150-fold molar excess. The reactions were subjected to electrophoresis in a 6% polyacrylamide gel at 180 V in 0.5X Tris/borate/ethylenediaminetetraacetic acid for ~1 h, and subsequently, the gel was dried and autoradiographed. (C) In vivo ChIP assay was performed for FOXA1 binding sites using an anti-FOXA1 antibody. The DNA elute from ChIP was subjected to polymerase chain reaction analysis from −321 to −464 and from −443 to −606 nucleotide positions for site 1 and site 2, respectively. FOXA1 , forkhead box protein A1; ERE, estrogen response element; ChIP, chromatin immunoprecipitation; EMSA, electrophoretic mobility shift assay; IP, immunoprecipitation; IgG, immunoglobulin G; Ntd, nucleotide; PS2 , trefoil factor 1.
Figure Legend Snippet: Schematic representation of the PS2 promoter. (A) Schematic diagram showing the presence of a functional estrogen response element (−407 nucleotide position) and two putative FOXA1 binding sites at −384 and −539 nucleotide positions, respectively. (B) In vitro binding assay. A total of 30 bp oligonucleotides containing FOXA1 binding sites were labeled with γ 32 P radioisotope and incubated with nuclear lysate extracted from MCF7 cells. An unlabeled FOXA1 (cold probe) consensus sequence was used for competition at 100 and 150-fold molar excess. The reactions were subjected to electrophoresis in a 6% polyacrylamide gel at 180 V in 0.5X Tris/borate/ethylenediaminetetraacetic acid for ~1 h, and subsequently, the gel was dried and autoradiographed. (C) In vivo ChIP assay was performed for FOXA1 binding sites using an anti-FOXA1 antibody. The DNA elute from ChIP was subjected to polymerase chain reaction analysis from −321 to −464 and from −443 to −606 nucleotide positions for site 1 and site 2, respectively. FOXA1 , forkhead box protein A1; ERE, estrogen response element; ChIP, chromatin immunoprecipitation; EMSA, electrophoretic mobility shift assay; IP, immunoprecipitation; IgG, immunoglobulin G; Ntd, nucleotide; PS2 , trefoil factor 1.

Techniques Used: Functional Assay, Binding Assay, In Vitro, Labeling, Incubation, Sequencing, Electrophoresis, In Vivo, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Electrophoretic Mobility Shift Assay, Immunoprecipitation

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    Millipore tris cl
    Release of glyoxal from the dihydro-dihydroxy form derived from <t>benznidazole.</t> (A) Schematic showing the release of glyoxal from dihydro-dihydroxyimidazole. (B) Spectrophotometric detection of glyoxal in reaction mixtures containing TbNTR (40 μg), benznidazole (200 μM), and various quantities of NADH (0 to 800 μM) in 1 ml 50 mM NaH 2 PO 4 (pH 7.5). To detect glyoxal, an aliquot of the reaction mixture was added to a borate buffer (pH 9.2) containing Girard T reagent, and the absorbance of the mixture at 325 nm was then determined. The concentration of glyoxal in the reaction mixtures was calculated by comparison against a standard curve obtained by using pure dialdehyde, after the subtraction of blank readings from samples lacking Girard T reagent. Values are means derived from experiments performed in triplicate ± standard deviations. (C) Time-dependent production of glyoxal from benznidazole. A 1-ml reaction mixture containing benznidazole (200 μM) in 50 mM <t>Tris-Cl</t> (pH 7.5) was incubated at room temperature with no addition (●), NADH (total amount added, 800 μM) (■), or NADH (total amount added, 800 μM) and 4 μg TbNTR (▴). At each time point, aliquots (100 μl) were taken and assayed for the presence of glyoxal. Glyoxal concentrations were then calculated as described above.
    Tris Cl, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tris cl/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tris cl - by Bioz Stars, 2022-01
    97/100 stars
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    Release of glyoxal from the dihydro-dihydroxy form derived from benznidazole. (A) Schematic showing the release of glyoxal from dihydro-dihydroxyimidazole. (B) Spectrophotometric detection of glyoxal in reaction mixtures containing TbNTR (40 μg), benznidazole (200 μM), and various quantities of NADH (0 to 800 μM) in 1 ml 50 mM NaH 2 PO 4 (pH 7.5). To detect glyoxal, an aliquot of the reaction mixture was added to a borate buffer (pH 9.2) containing Girard T reagent, and the absorbance of the mixture at 325 nm was then determined. The concentration of glyoxal in the reaction mixtures was calculated by comparison against a standard curve obtained by using pure dialdehyde, after the subtraction of blank readings from samples lacking Girard T reagent. Values are means derived from experiments performed in triplicate ± standard deviations. (C) Time-dependent production of glyoxal from benznidazole. A 1-ml reaction mixture containing benznidazole (200 μM) in 50 mM Tris-Cl (pH 7.5) was incubated at room temperature with no addition (●), NADH (total amount added, 800 μM) (■), or NADH (total amount added, 800 μM) and 4 μg TbNTR (▴). At each time point, aliquots (100 μl) were taken and assayed for the presence of glyoxal. Glyoxal concentrations were then calculated as described above.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Activation of Benznidazole by Trypanosomal Type I Nitroreductases Results in Glyoxal Formation

    doi: 10.1128/AAC.05135-11

    Figure Lengend Snippet: Release of glyoxal from the dihydro-dihydroxy form derived from benznidazole. (A) Schematic showing the release of glyoxal from dihydro-dihydroxyimidazole. (B) Spectrophotometric detection of glyoxal in reaction mixtures containing TbNTR (40 μg), benznidazole (200 μM), and various quantities of NADH (0 to 800 μM) in 1 ml 50 mM NaH 2 PO 4 (pH 7.5). To detect glyoxal, an aliquot of the reaction mixture was added to a borate buffer (pH 9.2) containing Girard T reagent, and the absorbance of the mixture at 325 nm was then determined. The concentration of glyoxal in the reaction mixtures was calculated by comparison against a standard curve obtained by using pure dialdehyde, after the subtraction of blank readings from samples lacking Girard T reagent. Values are means derived from experiments performed in triplicate ± standard deviations. (C) Time-dependent production of glyoxal from benznidazole. A 1-ml reaction mixture containing benznidazole (200 μM) in 50 mM Tris-Cl (pH 7.5) was incubated at room temperature with no addition (●), NADH (total amount added, 800 μM) (■), or NADH (total amount added, 800 μM) and 4 μg TbNTR (▴). At each time point, aliquots (100 μl) were taken and assayed for the presence of glyoxal. Glyoxal concentrations were then calculated as described above.

    Article Snippet: A standard assay mixture (200 μl) containing 50 mM Tris-Cl (pH 7.5), benznidazole (100 μM), glucose dehydrogenase (1 U ml−1 ), and glucose (3 mM) was equilibrated at 27°C for 15 min in the presence of TcNTR (4 μg) or cytochrome P450/cytochrome P450 reductase-enriched microsomal fractions (0.1 μg) (Sigma-Aldrich).

    Techniques: Derivative Assay, Concentration Assay, Incubation

    ∆48 PGAM5 forms tubular filaments in solution composed of ring-like structures. a Domain architecture of constructs used in this study. Full-length PGAM5 is comprised of a single transmembrane helix containing a mitochondrial targeting sequence (MTS), a linker domain including the regulatory multimerization motif (MM), and a C-terminal PGAM phosphatase domain. The native cleavage site between residues 24 and 25, cleaved by PARL 6 , is marked with a green arrow. b Elution profiles for ∆48 and ∆90 PGAM5 constructs purified by size exclusion chromatography (SEC) using a Superose 6 column (GE Healthcare) in SEC buffer containing 20 mM Tris-Cl pH 8.0, 500 µM TCEP, and 150 mM NaCl. The corresponding oligomeric states of each peak observed in the chromatograms are indicated. c Representative EM micrographs of negatively stained protein samples taken from fractions corresponding to the two distinct peaks observed in the ∆48 PGAM5 purification. d SEC profiles for ∆48 PGAM5 in SEC buffer and NaCl at a final concentration of 150, 300, or 750 mM. EM micrographs of negatively stained samples of ∆48 PGAM5 taken directly from the primary peak obtained during purification are shown, highlighting the decomposition of filaments into rings at increasing salt concentrations. Scale bars in c and d correspond to 50 nm, except for the inset in c in which the scale bar corresponds to 10 nm

    Journal: Nature Communications

    Article Title: Functional role of PGAM5 multimeric assemblies and their polymerization into filaments

    doi: 10.1038/s41467-019-08393-w

    Figure Lengend Snippet: ∆48 PGAM5 forms tubular filaments in solution composed of ring-like structures. a Domain architecture of constructs used in this study. Full-length PGAM5 is comprised of a single transmembrane helix containing a mitochondrial targeting sequence (MTS), a linker domain including the regulatory multimerization motif (MM), and a C-terminal PGAM phosphatase domain. The native cleavage site between residues 24 and 25, cleaved by PARL 6 , is marked with a green arrow. b Elution profiles for ∆48 and ∆90 PGAM5 constructs purified by size exclusion chromatography (SEC) using a Superose 6 column (GE Healthcare) in SEC buffer containing 20 mM Tris-Cl pH 8.0, 500 µM TCEP, and 150 mM NaCl. The corresponding oligomeric states of each peak observed in the chromatograms are indicated. c Representative EM micrographs of negatively stained protein samples taken from fractions corresponding to the two distinct peaks observed in the ∆48 PGAM5 purification. d SEC profiles for ∆48 PGAM5 in SEC buffer and NaCl at a final concentration of 150, 300, or 750 mM. EM micrographs of negatively stained samples of ∆48 PGAM5 taken directly from the primary peak obtained during purification are shown, highlighting the decomposition of filaments into rings at increasing salt concentrations. Scale bars in c and d correspond to 50 nm, except for the inset in c in which the scale bar corresponds to 10 nm

    Article Snippet: Protein was eluted using Elution Buffer (20 mM Tris-Cl pH 8.0, 500 mM NaCl, 0.5 mM TCEP, 5% glycerol, 200 mM Imidazole pH 8.0) and exchanged into buffer containing 20 mM Tris-Cl pH 8.0, 150 mM NaCl, and 0.5 mM TCEP using a 10 kDa cutoff concentrator (Millipore).

    Techniques: Construct, Sequencing, Purification, Size-exclusion Chromatography, Staining, Concentration Assay

    Size exclusion chromatography profiles upon separation using Superdex 200 column (GE Healthcare). A , standard gel filtration marker (Bio-Rad catalog number 151-1901). B , Ec ASNase1 chromatogram in 50 m m Tris-Cl, 100 m m NaCl, pH 8. C , profile of hASNase1

    Journal: The Journal of Biological Chemistry

    Article Title: Human 60-kDa Lysophospholipase Contains an N-terminal l-Asparaginase Domain That Is Allosterically Regulated by l-Asparagine *

    doi: 10.1074/jbc.M113.545038

    Figure Lengend Snippet: Size exclusion chromatography profiles upon separation using Superdex 200 column (GE Healthcare). A , standard gel filtration marker (Bio-Rad catalog number 151-1901). B , Ec ASNase1 chromatogram in 50 m m Tris-Cl, 100 m m NaCl, pH 8. C , profile of hASNase1

    Article Snippet: Enzyme samples were dialyzed (Slide-A-Lyzer, Pierce; 10,000 molecular weight cutoff) against 50 m m Tris-Cl, 0.1 NaCl, pH 8 to remove glycerol and subsequently mixed with SYPRO Orange (Sigma-Aldrich) in a final volume of 20 μl.

    Techniques: Size-exclusion Chromatography, Filtration, Marker

    Effect of pH on hASNase1 activity ( A ) and stability ( B ). Buffers used were: sodium acetate (pH 3–5), sodium phosphate (pH 6–7), Tris-Cl (pH 7–8.5), and CAPSO (pH 9–10), all at 50 m m concentration in 100 m m NaCl. Enzymatic

    Journal: The Journal of Biological Chemistry

    Article Title: Human 60-kDa Lysophospholipase Contains an N-terminal l-Asparaginase Domain That Is Allosterically Regulated by l-Asparagine *

    doi: 10.1074/jbc.M113.545038

    Figure Lengend Snippet: Effect of pH on hASNase1 activity ( A ) and stability ( B ). Buffers used were: sodium acetate (pH 3–5), sodium phosphate (pH 6–7), Tris-Cl (pH 7–8.5), and CAPSO (pH 9–10), all at 50 m m concentration in 100 m m NaCl. Enzymatic

    Article Snippet: Enzyme samples were dialyzed (Slide-A-Lyzer, Pierce; 10,000 molecular weight cutoff) against 50 m m Tris-Cl, 0.1 NaCl, pH 8 to remove glycerol and subsequently mixed with SYPRO Orange (Sigma-Aldrich) in a final volume of 20 μl.

    Techniques: Activity Assay, Concentration Assay

    Substrate specificity of picNuA4. All reactions contained 1 × TBA at pH 7.0 or 50 mM Tris at pH 7.2, 1 mM DTT, 50–100 μ M acetyl-CoA, and 0.01–0.1 μ M picNuA4 complex. Concentrations of peptide substrates used in saturation curves were varied over a 10-fold range covering concentrations above and below the K m . Data were fitted to appropriate equations in Kaleidagraph to determine the kinetic constants. Each value is an average of three to six separate assays, with the standard deviation shown. Gray bars represent values from saturation curves ( k cat / K m ), and black bars represent values determined from progress curves ( k cat / K avg ). (***) Value represents the lower limit for initial k cat / K m of NCPs because data below 150 nM were not collected.

    Journal: Biochemistry

    Article Title: Nucleosome Recognition by the Piccolo NuA4 Histone Acetyltransferase Complex †

    doi: 10.1021/bi602366n

    Figure Lengend Snippet: Substrate specificity of picNuA4. All reactions contained 1 × TBA at pH 7.0 or 50 mM Tris at pH 7.2, 1 mM DTT, 50–100 μ M acetyl-CoA, and 0.01–0.1 μ M picNuA4 complex. Concentrations of peptide substrates used in saturation curves were varied over a 10-fold range covering concentrations above and below the K m . Data were fitted to appropriate equations in Kaleidagraph to determine the kinetic constants. Each value is an average of three to six separate assays, with the standard deviation shown. Gray bars represent values from saturation curves ( k cat / K m ), and black bars represent values determined from progress curves ( k cat / K avg ). (***) Value represents the lower limit for initial k cat / K m of NCPs because data below 150 nM were not collected.

    Article Snippet: Acetyl-CoA, Tris-Cl, Bis-tris, sodium acetate, dithiothreitol (DTT), and other reagents were purchased from Sigma-Aldrich or Fisher Chemicals.

    Techniques: Standard Deviation

    Substrate specificity of Esa1. All reactions contain 1 × TBA at pH 7.0 or 50 mM Tris at pH 7.2, 1 mM DTT, 75–100 μ M acetyl-CoA, and 0.54–2 μ M Esa1. Concentrations of peptide substrates in saturation curves were varied over a 10-fold range covering concentrations above and below the K m . Data were fitted to the Michaelis–Menten equation in Kaleidagraph to determine kinetic constants. The k cat / K m value for nucleosome arrays was collected under subsaturating conditions, and accordingly, the data were fitted to a linear equation where the slope is k cat / K m.

    Journal: Biochemistry

    Article Title: Nucleosome Recognition by the Piccolo NuA4 Histone Acetyltransferase Complex †

    doi: 10.1021/bi602366n

    Figure Lengend Snippet: Substrate specificity of Esa1. All reactions contain 1 × TBA at pH 7.0 or 50 mM Tris at pH 7.2, 1 mM DTT, 75–100 μ M acetyl-CoA, and 0.54–2 μ M Esa1. Concentrations of peptide substrates in saturation curves were varied over a 10-fold range covering concentrations above and below the K m . Data were fitted to the Michaelis–Menten equation in Kaleidagraph to determine kinetic constants. The k cat / K m value for nucleosome arrays was collected under subsaturating conditions, and accordingly, the data were fitted to a linear equation where the slope is k cat / K m.

    Article Snippet: Acetyl-CoA, Tris-Cl, Bis-tris, sodium acetate, dithiothreitol (DTT), and other reagents were purchased from Sigma-Aldrich or Fisher Chemicals.

    Techniques: