cardiomyocytes  (Worthington Biochemical)


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    Structured Review

    Worthington Biochemical cardiomyocytes
    SUMO-1 overexpression protects SERCA2a activity from oxidative stress. (A) <t>Cardiomyocytes</t> were infected with adenoviruses carrying SUMO-1 or β-gal genes for 24 h and then, the cells were exposed to 0.5 μ M of H 2 O 2 for 6 h. SERCA2a SUMOylation was determined by IP with anti-SUMO-1 followed by WB with anti-SERCA2a antibody. Tyrosine nitration of SERCA2a was measured by IP with anti-SERCA2a followed by WB with anti-3-nitrotyrosine antibody. The cell extracts were subjected to ATPase activity assay of SERCA2a. (B) Cardiac-specific Nox4 knockout mice were subjected to TAC operation for 4 weeks. The expression levels of SERCA2a, SUMO-1, and SUMOylated SERCA2a in cardiac tissues of Nox4 knockout mice are shown. Data indicate the mean±SD of three independent experiments. * p
    Cardiomyocytes, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "The Role of SUMO-1 in Cardiac Oxidative Stress and Hypertrophy"

    Article Title: The Role of SUMO-1 in Cardiac Oxidative Stress and Hypertrophy

    Journal: Antioxidants & Redox Signaling

    doi: 10.1089/ars.2014.5983

    SUMO-1 overexpression protects SERCA2a activity from oxidative stress. (A) Cardiomyocytes were infected with adenoviruses carrying SUMO-1 or β-gal genes for 24 h and then, the cells were exposed to 0.5 μ M of H 2 O 2 for 6 h. SERCA2a SUMOylation was determined by IP with anti-SUMO-1 followed by WB with anti-SERCA2a antibody. Tyrosine nitration of SERCA2a was measured by IP with anti-SERCA2a followed by WB with anti-3-nitrotyrosine antibody. The cell extracts were subjected to ATPase activity assay of SERCA2a. (B) Cardiac-specific Nox4 knockout mice were subjected to TAC operation for 4 weeks. The expression levels of SERCA2a, SUMO-1, and SUMOylated SERCA2a in cardiac tissues of Nox4 knockout mice are shown. Data indicate the mean±SD of three independent experiments. * p
    Figure Legend Snippet: SUMO-1 overexpression protects SERCA2a activity from oxidative stress. (A) Cardiomyocytes were infected with adenoviruses carrying SUMO-1 or β-gal genes for 24 h and then, the cells were exposed to 0.5 μ M of H 2 O 2 for 6 h. SERCA2a SUMOylation was determined by IP with anti-SUMO-1 followed by WB with anti-SERCA2a antibody. Tyrosine nitration of SERCA2a was measured by IP with anti-SERCA2a followed by WB with anti-3-nitrotyrosine antibody. The cell extracts were subjected to ATPase activity assay of SERCA2a. (B) Cardiac-specific Nox4 knockout mice were subjected to TAC operation for 4 weeks. The expression levels of SERCA2a, SUMO-1, and SUMOylated SERCA2a in cardiac tissues of Nox4 knockout mice are shown. Data indicate the mean±SD of three independent experiments. * p

    Techniques Used: Over Expression, Activity Assay, Infection, Western Blot, Nitration, Knock-Out, Mouse Assay, Expressing

    SUMO-1 overexpression inhibits PE-induced hypertrophic response in cardiomyocytes. (A) The neonatal cardiomyocytes were infected with the indicated adenoviruses and further stimulated with PE (100 μ M ) for 24 h. The relative expression levels of hypertrophic association genes such as ANF, BNP, and myh7 were determined by semi-quantitative RT-PCR ( n =3). (B) The protein synthesis rates after PE stimulation were evaluated by measuring 3 H-leucine incorporation. Data indicate the mean value±SD of three independent experiments. (C) Cell size measurement. (D) MAPK activities after the stimulation of neonatal cardiomyocytes with PE. Cardiomyocytes were infected with indicated adenoviruses for 24 h and then treated with PE (100 μ M ) for 24 h. Cell extracts were examined for amounts of phosphorylated ERK, p38, JNK, and c-Jun using phospho-specific antibodies. Total protein levels of ERK, p38, JNK, and c-Jun were also analyzed. The relative intensity was quantitated using ImageJ software. Data indicate the mean±SD of three independent experiments. # p
    Figure Legend Snippet: SUMO-1 overexpression inhibits PE-induced hypertrophic response in cardiomyocytes. (A) The neonatal cardiomyocytes were infected with the indicated adenoviruses and further stimulated with PE (100 μ M ) for 24 h. The relative expression levels of hypertrophic association genes such as ANF, BNP, and myh7 were determined by semi-quantitative RT-PCR ( n =3). (B) The protein synthesis rates after PE stimulation were evaluated by measuring 3 H-leucine incorporation. Data indicate the mean value±SD of three independent experiments. (C) Cell size measurement. (D) MAPK activities after the stimulation of neonatal cardiomyocytes with PE. Cardiomyocytes were infected with indicated adenoviruses for 24 h and then treated with PE (100 μ M ) for 24 h. Cell extracts were examined for amounts of phosphorylated ERK, p38, JNK, and c-Jun using phospho-specific antibodies. Total protein levels of ERK, p38, JNK, and c-Jun were also analyzed. The relative intensity was quantitated using ImageJ software. Data indicate the mean±SD of three independent experiments. # p

    Techniques Used: Over Expression, Infection, Expressing, Quantitative RT-PCR, Software

    Related Articles

    other:

    Article Title: Multifaceted roles of miR-1s in repressing the fetal gene program in the heart
    Article Snippet: Briefly, neonatal hearts at P2.5 were dissected out and minced into small pieces, and cardiomyocytes were released by digestion with 229 U/ml collagenase type II (Worthington Biochemical).

    Isolation:

    Article Title: Mobilized bone marrow progenitor cells serve as donors of cytoprotective genes for cardiac repair
    Article Snippet: .. To demonstrate the direct protection of MPCs on cardiomyocytes, MPCs were co-cultured with cardiomyocytes which were isolated from ventricles of 2-day-old neonatal Sprague-Dawley rats (Harlan, Indianapolis, IN) using the Neonatal Cardiomyocyte Isolation System (Worthington biochemical Co. NJ) as previously described ( ). ..

    Article Title: Heat‐shock transcription factor 1 is critically involved in the ischaemia‐induced cardiac hypertrophy via JAK2/ STAT3 pathway, et al. Heat‐shock transcription factor 1 is critically involved in the ischaemia‐induced cardiac hypertrophy via JAK2/STAT3 pathway
    Article Snippet: .. Enzymatic and mechanical dissociation of cardiomyocytes was then performed using the Neonatal Cardiomyocyte Isolation System supplied by Worthington Biochemicals (USA). ..

    Article Title: Improvement of cardiomyocyte function by in vivo hexarelin treatment in streptozotocin‐induced diabetic rats. Improvement of cardiomyocyte function by in vivo hexarelin treatment in streptozotocin‐induced diabetic rats
    Article Snippet: .. Following this, cardiomyocytes were isolated from the left ventricle of each heart with Tyrode solution containing 100 μ mol/L CaCl2 , 0.5 mg/mL collagenase Type II (Worthington, NJ, USA) and 0.15 mg/mL proteinase Type XXIV (Sigma, MO, USA). ..

    Article Title: FGF23 is a novel regulator of intracellular calcium and cardiac contractility in addition to cardiac hypertrophy
    Article Snippet: .. Cardiomyocytes were isolated in a standard manner, utilizing retrograde perfusion via a proprietary procedure developed in our laboratory with Worthington Biochemical (Lakewood, NJ). ..

    Article Title: The Role of SUMO-1 in Cardiac Oxidative Stress and Hypertrophy
    Article Snippet: .. Briefly, ventricular tissue was enzymatically dissociated, and the resulting cardiomyocytes was isolated using Neonatal Cardiomyocyte isolation kit (Worthington Biochemical). ..

    Article Title: Overexpression of miR-223 Tips the Balance of Pro- and Anti-hypertrophic Signaling Cascades toward Physiologic Cardiac Hypertrophy *
    Article Snippet: .. Hearts of rat neonates (1–3 days old) were removed after anesthetizing by ice-water bath, and cardiomyocytes were isolated using the Worthington Neonatal Cardiomyocytes Isolation System (Worthington Biochemical Corp., Lakewood, NJ) following the manufacturer's protocol. ..

    Western Blot:

    Article Title: Extracellular vesicles derived from microRNA-150-5p-overexpressing mesenchymal stem cells protect rat hearts against ischemia/reperfusion
    Article Snippet: .. Western blot analysisTotal protein was extracted from myocardium and cardiomyocytes in the ischemic border area and quantified by BCA kit (Beyotime, Jiangsu, China), followed by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. ..

    Nucleic Acid Electrophoresis:

    Article Title: Extracellular vesicles derived from microRNA-150-5p-overexpressing mesenchymal stem cells protect rat hearts against ischemia/reperfusion
    Article Snippet: .. Western blot analysisTotal protein was extracted from myocardium and cardiomyocytes in the ischemic border area and quantified by BCA kit (Beyotime, Jiangsu, China), followed by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. ..

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    Worthington Biochemical fluorescent activated cell sorting neurospheres
    Neurosphere differentiation and composition. ( A and B ) 10 days-post ES cell differentiation <t>neurospheres</t> were dissociated and cell composition was quantified using FACS. Plots of V3, V2a, and V1 interneurons and motor neurons from Sim1:Cre;R26/C:LSL:Tomato, Chx10:Cre;R26/C:LSL:Tomato, En1:Cre;R26/C:LSL:Tomato, and Hb9:GFP ES cell reporter lines, respectively are shown after differentiation with ( A ) 1000 nM and ( B ) 5 nM SAG. ( C ) Two Chx:10:Cre;R26/C:LSL:Tomato ES cell lines were differentiated 10 days under a range of SAG concentrations. Tomato+V2a neuron number was quantified using FACS. ( D ) Chx:10:Cre;R26/C:LSL:Tomato lines #1 and #2 were differentiated with 15 nM and 40 nM SAG, respectively. 5 μM DAPT was applied days 6–10. Standardized tomato + V2a neurons relative to no DAPT control. DOI: http://dx.doi.org/10.7554/eLife.21540.004
    Fluorescent Activated Cell Sorting Neurospheres, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Worthington Biochemical α amylase inhibitory activity
    <t>α‐Amylase</t> inhibitory activities of butternut squash residues (seed and skin extract). Values are means of duplicate determinations
    α Amylase Inhibitory Activity, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Worthington Biochemical fret activity assays nuc enzyme
    Activity of <t>Nuc/Nuc2</t> chimeric proteins. Chimeras consisting of combinations of Nuc and Nuc2 N- and C-terminal protein domains were constructed. Each chimera was constructed to include a His 6 -tag on the C-terminal protein domain. A . Schematic showing the Nuc/Nuc2 chimeric proteins constructed, with Nuc domains colored in teal and Nuc2 domains colored in purple (see Fig. 1A for additional description of domains and processing sites). Chimeras constructed include, from the top down: Nuc control (pMK12), Nuc2 control (pMK13), Nuc signal sequence fused to the Nuc2 active domain (pMK8), Nuc signal sequence and NucB leader fused to the Nuc2 active domain (pMK9), Nuc2 membrane anchor fused to NucA (pMK10) and Nuc2 membrane anchor fused to NucB (pMK11). Each construct was expressed in a S. aureus nuc mutant (strain AH1680) for 15 hr, and culture supernatant was saved and membranes purified. Anti-His immunoblots were performed to detect the presence of C-terminal His 6 -tag on chimeric proteins in culture supernatants ( B ) or membrane ( C ) fractions. <t>FRET</t> activity assays were performed to detect nuclease activity in culture supernatants ( D ) and membrane ( E) fractions. The nuc mutant strain was included as a negative control (called “no plasmid”).
    Fret Activity Assays Nuc Enzyme, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Worthington Biochemical dnase i activity
    Testing the inhibition of deoxyribonuclease I (DNase I) by previously known in vitro inhibitors. To generate dose–response curves, the change in product/min (pmole/min) by <t>DNase</t> I (1.72 nM) was monitored throughout a series of concentrations for
    Dnase I Activity, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Neurosphere differentiation and composition. ( A and B ) 10 days-post ES cell differentiation neurospheres were dissociated and cell composition was quantified using FACS. Plots of V3, V2a, and V1 interneurons and motor neurons from Sim1:Cre;R26/C:LSL:Tomato, Chx10:Cre;R26/C:LSL:Tomato, En1:Cre;R26/C:LSL:Tomato, and Hb9:GFP ES cell reporter lines, respectively are shown after differentiation with ( A ) 1000 nM and ( B ) 5 nM SAG. ( C ) Two Chx:10:Cre;R26/C:LSL:Tomato ES cell lines were differentiated 10 days under a range of SAG concentrations. Tomato+V2a neuron number was quantified using FACS. ( D ) Chx:10:Cre;R26/C:LSL:Tomato lines #1 and #2 were differentiated with 15 nM and 40 nM SAG, respectively. 5 μM DAPT was applied days 6–10. Standardized tomato + V2a neurons relative to no DAPT control. DOI: http://dx.doi.org/10.7554/eLife.21540.004

    Journal: eLife

    Article Title: Speed and segmentation control mechanisms characterized in rhythmically-active circuits created from spinal neurons produced from genetically-tagged embryonic stem cells

    doi: 10.7554/eLife.21540

    Figure Lengend Snippet: Neurosphere differentiation and composition. ( A and B ) 10 days-post ES cell differentiation neurospheres were dissociated and cell composition was quantified using FACS. Plots of V3, V2a, and V1 interneurons and motor neurons from Sim1:Cre;R26/C:LSL:Tomato, Chx10:Cre;R26/C:LSL:Tomato, En1:Cre;R26/C:LSL:Tomato, and Hb9:GFP ES cell reporter lines, respectively are shown after differentiation with ( A ) 1000 nM and ( B ) 5 nM SAG. ( C ) Two Chx:10:Cre;R26/C:LSL:Tomato ES cell lines were differentiated 10 days under a range of SAG concentrations. Tomato+V2a neuron number was quantified using FACS. ( D ) Chx:10:Cre;R26/C:LSL:Tomato lines #1 and #2 were differentiated with 15 nM and 40 nM SAG, respectively. 5 μM DAPT was applied days 6–10. Standardized tomato + V2a neurons relative to no DAPT control. DOI: http://dx.doi.org/10.7554/eLife.21540.004

    Article Snippet: Fluorescent-activated cell sorting Neurospheres, 6–11 days from ES cells, were dissociated (Papain, Worthington), and then counted with a BD FACScan to determine the percentage of neurons composing neurospheres at different SAG concentrations.

    Techniques: Cell Differentiation, FACS

    α‐Amylase inhibitory activities of butternut squash residues (seed and skin extract). Values are means of duplicate determinations

    Journal: Food Science & Nutrition

    Article Title: Evaluation of bioactivity of butternut squash (Cucurbita moschata D.) seeds and skin. Evaluation of bioactivity of butternut squash (Cucurbita moschata D.) seeds and skin

    doi: 10.1002/fsn3.1602

    Figure Lengend Snippet: α‐Amylase inhibitory activities of butternut squash residues (seed and skin extract). Values are means of duplicate determinations

    Article Snippet: The α‐amylase inhibitory activity was expressed as percentage inhibition (Worthington, Weddell, & Neilson, ).

    Techniques:

    Activity of Nuc/Nuc2 chimeric proteins. Chimeras consisting of combinations of Nuc and Nuc2 N- and C-terminal protein domains were constructed. Each chimera was constructed to include a His 6 -tag on the C-terminal protein domain. A . Schematic showing the Nuc/Nuc2 chimeric proteins constructed, with Nuc domains colored in teal and Nuc2 domains colored in purple (see Fig. 1A for additional description of domains and processing sites). Chimeras constructed include, from the top down: Nuc control (pMK12), Nuc2 control (pMK13), Nuc signal sequence fused to the Nuc2 active domain (pMK8), Nuc signal sequence and NucB leader fused to the Nuc2 active domain (pMK9), Nuc2 membrane anchor fused to NucA (pMK10) and Nuc2 membrane anchor fused to NucB (pMK11). Each construct was expressed in a S. aureus nuc mutant (strain AH1680) for 15 hr, and culture supernatant was saved and membranes purified. Anti-His immunoblots were performed to detect the presence of C-terminal His 6 -tag on chimeric proteins in culture supernatants ( B ) or membrane ( C ) fractions. FRET activity assays were performed to detect nuclease activity in culture supernatants ( D ) and membrane ( E) fractions. The nuc mutant strain was included as a negative control (called “no plasmid”).

    Journal: PLoS ONE

    Article Title: Staphylococcus aureus Nuc2 Is a Functional, Surface-Attached Extracellular Nuclease

    doi: 10.1371/journal.pone.0095574

    Figure Lengend Snippet: Activity of Nuc/Nuc2 chimeric proteins. Chimeras consisting of combinations of Nuc and Nuc2 N- and C-terminal protein domains were constructed. Each chimera was constructed to include a His 6 -tag on the C-terminal protein domain. A . Schematic showing the Nuc/Nuc2 chimeric proteins constructed, with Nuc domains colored in teal and Nuc2 domains colored in purple (see Fig. 1A for additional description of domains and processing sites). Chimeras constructed include, from the top down: Nuc control (pMK12), Nuc2 control (pMK13), Nuc signal sequence fused to the Nuc2 active domain (pMK8), Nuc signal sequence and NucB leader fused to the Nuc2 active domain (pMK9), Nuc2 membrane anchor fused to NucA (pMK10) and Nuc2 membrane anchor fused to NucB (pMK11). Each construct was expressed in a S. aureus nuc mutant (strain AH1680) for 15 hr, and culture supernatant was saved and membranes purified. Anti-His immunoblots were performed to detect the presence of C-terminal His 6 -tag on chimeric proteins in culture supernatants ( B ) or membrane ( C ) fractions. FRET activity assays were performed to detect nuclease activity in culture supernatants ( D ) and membrane ( E) fractions. The nuc mutant strain was included as a negative control (called “no plasmid”).

    Article Snippet: FRET activity assays Nuc enzyme was purchased from Worthington Biochemical (Lakewood, NJ).

    Techniques: Activity Assay, Construct, Sequencing, Mutagenesis, Purification, Western Blot, Negative Control, Plasmid Preparation

    Nuc2 is expressed at low levels during growth. A . Monitoring changes in fluorescence of a P nuc2 -sGFP transcriptional reporter over time (open circles) in comparison to growth (black squares). Dotted line indicates level of background fluorescence of growth media. B . Changes in surface Nuc2 activity (open circles) present in a S. aureus USA300 LAC nuc mutant over time as measured by whole-cell FRET assay.

    Journal: PLoS ONE

    Article Title: Staphylococcus aureus Nuc2 Is a Functional, Surface-Attached Extracellular Nuclease

    doi: 10.1371/journal.pone.0095574

    Figure Lengend Snippet: Nuc2 is expressed at low levels during growth. A . Monitoring changes in fluorescence of a P nuc2 -sGFP transcriptional reporter over time (open circles) in comparison to growth (black squares). Dotted line indicates level of background fluorescence of growth media. B . Changes in surface Nuc2 activity (open circles) present in a S. aureus USA300 LAC nuc mutant over time as measured by whole-cell FRET assay.

    Article Snippet: FRET activity assays Nuc enzyme was purchased from Worthington Biochemical (Lakewood, NJ).

    Techniques: Fluorescence, Activity Assay, Mutagenesis

    Nuc2 activity can be measured at the cell surface. A . Whole-cell FRET activity assay measuring Nuc2 surface function with UAMS-1 WT, nuc2 , nuc and nuc nuc2 double mutants. B . Cell surface nuclease activity in USA300 strain LAC WT, nuc2 , nuc and nuc nuc2 double mutants. Statistical significance (*p

    Journal: PLoS ONE

    Article Title: Staphylococcus aureus Nuc2 Is a Functional, Surface-Attached Extracellular Nuclease

    doi: 10.1371/journal.pone.0095574

    Figure Lengend Snippet: Nuc2 activity can be measured at the cell surface. A . Whole-cell FRET activity assay measuring Nuc2 surface function with UAMS-1 WT, nuc2 , nuc and nuc nuc2 double mutants. B . Cell surface nuclease activity in USA300 strain LAC WT, nuc2 , nuc and nuc nuc2 double mutants. Statistical significance (*p

    Article Snippet: FRET activity assays Nuc enzyme was purchased from Worthington Biochemical (Lakewood, NJ).

    Techniques: Activity Assay

    Testing the inhibition of deoxyribonuclease I (DNase I) by previously known in vitro inhibitors. To generate dose–response curves, the change in product/min (pmole/min) by DNase I (1.72 nM) was monitored throughout a series of concentrations for

    Journal: Journal of biomolecular screening

    Article Title: Novel High-Throughput Deoxyribonuclease 1 Assay

    doi: 10.1177/1087057114555828

    Figure Lengend Snippet: Testing the inhibition of deoxyribonuclease I (DNase I) by previously known in vitro inhibitors. To generate dose–response curves, the change in product/min (pmole/min) by DNase I (1.72 nM) was monitored throughout a series of concentrations for

    Article Snippet: The percentage of DNase I activity was calculated using Equation 1: DNase\u00a0I\u00a0activity (%) =\u00a0 (mean\u00a0velocity\u00a0of\u00a0a\u00a0compound/mean\u00a0velocity\u00a0of\u00a0DMSO)\u00a0\u00d7\u00a0100 (1) In similar assays, recombinant murine EndoG (produced in-house) was used at a concentration of 0.14 μM in 0.1 mM MgCl2 , 10 mM Tris-HCl, pH 7.4; and DNase II (Worthington, Lakewood, NJ) (3.32 nM) was tested in 100 mM sodium citrate buffer, pH 5.0.

    Techniques: Inhibition, In Vitro

    Specificity of the high-throughput screening (HTS) assay to deoxyribonuclease I (DNase I). A Michaelis–Menten curve for each of 3 endonucleases (1.72 nM each) was generated for the assay to afford kinetic analysis. k cat / K m values of DNase I and

    Journal: Journal of biomolecular screening

    Article Title: Novel High-Throughput Deoxyribonuclease 1 Assay

    doi: 10.1177/1087057114555828

    Figure Lengend Snippet: Specificity of the high-throughput screening (HTS) assay to deoxyribonuclease I (DNase I). A Michaelis–Menten curve for each of 3 endonucleases (1.72 nM each) was generated for the assay to afford kinetic analysis. k cat / K m values of DNase I and

    Article Snippet: The percentage of DNase I activity was calculated using Equation 1: DNase\u00a0I\u00a0activity (%) =\u00a0 (mean\u00a0velocity\u00a0of\u00a0a\u00a0compound/mean\u00a0velocity\u00a0of\u00a0DMSO)\u00a0\u00d7\u00a0100 (1) In similar assays, recombinant murine EndoG (produced in-house) was used at a concentration of 0.14 μM in 0.1 mM MgCl2 , 10 mM Tris-HCl, pH 7.4; and DNase II (Worthington, Lakewood, NJ) (3.32 nM) was tested in 100 mM sodium citrate buffer, pH 5.0.

    Techniques: High Throughput Screening Assay, HTS Assay, Generated

    Identification of deoxyribonuclease I (DNase I) inhibitor candidates by screening of the chemical library of 1040 compounds. The library was screened at ( A ) 10 μM and ( B ) 1 μM concentrations, affording 45 and 5 hits, respectively. The

    Journal: Journal of biomolecular screening

    Article Title: Novel High-Throughput Deoxyribonuclease 1 Assay

    doi: 10.1177/1087057114555828

    Figure Lengend Snippet: Identification of deoxyribonuclease I (DNase I) inhibitor candidates by screening of the chemical library of 1040 compounds. The library was screened at ( A ) 10 μM and ( B ) 1 μM concentrations, affording 45 and 5 hits, respectively. The

    Article Snippet: The percentage of DNase I activity was calculated using Equation 1: DNase\u00a0I\u00a0activity (%) =\u00a0 (mean\u00a0velocity\u00a0of\u00a0a\u00a0compound/mean\u00a0velocity\u00a0of\u00a0DMSO)\u00a0\u00d7\u00a0100 (1) In similar assays, recombinant murine EndoG (produced in-house) was used at a concentration of 0.14 μM in 0.1 mM MgCl2 , 10 mM Tris-HCl, pH 7.4; and DNase II (Worthington, Lakewood, NJ) (3.32 nM) was tested in 100 mM sodium citrate buffer, pH 5.0.

    Techniques:

    Elaboration of the high-throughput screening (HTS) deoxyribonuclease I (DNase I) assay conditions. ( A ) A schematic diagram for the substrate, Cy5.5-end-labeled oligonucleotide, and principle of the assay. ( B ) The effects of the concentrations of calcium,

    Journal: Journal of biomolecular screening

    Article Title: Novel High-Throughput Deoxyribonuclease 1 Assay

    doi: 10.1177/1087057114555828

    Figure Lengend Snippet: Elaboration of the high-throughput screening (HTS) deoxyribonuclease I (DNase I) assay conditions. ( A ) A schematic diagram for the substrate, Cy5.5-end-labeled oligonucleotide, and principle of the assay. ( B ) The effects of the concentrations of calcium,

    Article Snippet: The percentage of DNase I activity was calculated using Equation 1: DNase\u00a0I\u00a0activity (%) =\u00a0 (mean\u00a0velocity\u00a0of\u00a0a\u00a0compound/mean\u00a0velocity\u00a0of\u00a0DMSO)\u00a0\u00d7\u00a0100 (1) In similar assays, recombinant murine EndoG (produced in-house) was used at a concentration of 0.14 μM in 0.1 mM MgCl2 , 10 mM Tris-HCl, pH 7.4; and DNase II (Worthington, Lakewood, NJ) (3.32 nM) was tested in 100 mM sodium citrate buffer, pH 5.0.

    Techniques: High Throughput Screening Assay, Labeling