cardiomyocytes  (Worthington Biochemical)

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

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:

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

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

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

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

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

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:

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