Incubation:Article Title: Early neuronal accumulation of DNA double strand breaks in Alzheimer’s disease
Article Snippet: Briefly, sections were washed in Tris-buffered saline (TBS), permeabilized in TBS containing 0.5% Triton X-100 (Sigma-Aldrich) at 4 °C overnight, and incubated for 15 min at 110 °C in citrate buffer (pH 6.0) for antigen retrieval. .. They were then incubated in 3% H2 O2 and 10% methanol for 15 min, 0.1% Sudan Black (Sigma-Aldrich) in 70% ethanol for 15 min, and 10% goat serum in TBS containing 0.1% Tween-20 (TBST) for 1 h. Anti-γH2AX and anti-NeuN antibodies (Additional file : Table S3) in TBST containing 3% goat serum were applied first at 4 °C overnight and then at room temperature for 2 h. Primary antibodies were detected with Alexa Fluor secondary antibodies (Thermo Fisher Scientific) at 1:500 dilution. ..
Staining:Article Title: Penfluridol triggers cytoprotective autophagy and cellular apoptosis through ROS induction and activation of the PP2A-modulated MAPK pathway in acute myeloid leukemia with different FLT3 statuses
Article Snippet: Densitometric analysis of the bands was carried out using Image-Pro Plus software (Media Cybernetics, Rockville, MD). .. Measurement of reactive oxygen species (ROS) production AML cells were treated with penfluridol or H2 O2 for the indicated time points, and the intracellular ROS production was detected by staining with the fluoroprobe, 2′,7′-dichlorofluorescin diacetate (DCFDA, Sigma), in RPMI 1640 medium for 30 min. After washing cells with PBS, ROS production of DCFDA-preloaded cells was captured using a fluorescence microscope (Zeiss Axioplan) or quantified by flow cytometry. .. Detection and quantification of autophagic cells by staining with acridine orange (AO) Formation of acidic vesicular organelles (AVOs), a morphological characteristic of autophagy, was detected by AO (Sigma) staining.
Fluorescence:Article Title: Penfluridol triggers cytoprotective autophagy and cellular apoptosis through ROS induction and activation of the PP2A-modulated MAPK pathway in acute myeloid leukemia with different FLT3 statuses
Article Snippet: Densitometric analysis of the bands was carried out using Image-Pro Plus software (Media Cybernetics, Rockville, MD). .. Measurement of reactive oxygen species (ROS) production AML cells were treated with penfluridol or H2 O2 for the indicated time points, and the intracellular ROS production was detected by staining with the fluoroprobe, 2′,7′-dichlorofluorescin diacetate (DCFDA, Sigma), in RPMI 1640 medium for 30 min. After washing cells with PBS, ROS production of DCFDA-preloaded cells was captured using a fluorescence microscope (Zeiss Axioplan) or quantified by flow cytometry. .. Detection and quantification of autophagic cells by staining with acridine orange (AO) Formation of acidic vesicular organelles (AVOs), a morphological characteristic of autophagy, was detected by AO (Sigma) staining.
Microscopy:Article Title: Penfluridol triggers cytoprotective autophagy and cellular apoptosis through ROS induction and activation of the PP2A-modulated MAPK pathway in acute myeloid leukemia with different FLT3 statuses
Article Snippet: Densitometric analysis of the bands was carried out using Image-Pro Plus software (Media Cybernetics, Rockville, MD). .. Measurement of reactive oxygen species (ROS) production AML cells were treated with penfluridol or H2 O2 for the indicated time points, and the intracellular ROS production was detected by staining with the fluoroprobe, 2′,7′-dichlorofluorescin diacetate (DCFDA, Sigma), in RPMI 1640 medium for 30 min. After washing cells with PBS, ROS production of DCFDA-preloaded cells was captured using a fluorescence microscope (Zeiss Axioplan) or quantified by flow cytometry. .. Detection and quantification of autophagic cells by staining with acridine orange (AO) Formation of acidic vesicular organelles (AVOs), a morphological characteristic of autophagy, was detected by AO (Sigma) staining.
Flow Cytometry:Article Title: Penfluridol triggers cytoprotective autophagy and cellular apoptosis through ROS induction and activation of the PP2A-modulated MAPK pathway in acute myeloid leukemia with different FLT3 statuses
Article Snippet: Densitometric analysis of the bands was carried out using Image-Pro Plus software (Media Cybernetics, Rockville, MD). .. Measurement of reactive oxygen species (ROS) production AML cells were treated with penfluridol or H2 O2 for the indicated time points, and the intracellular ROS production was detected by staining with the fluoroprobe, 2′,7′-dichlorofluorescin diacetate (DCFDA, Sigma), in RPMI 1640 medium for 30 min. After washing cells with PBS, ROS production of DCFDA-preloaded cells was captured using a fluorescence microscope (Zeiss Axioplan) or quantified by flow cytometry. .. Detection and quantification of autophagic cells by staining with acridine orange (AO) Formation of acidic vesicular organelles (AVOs), a morphological characteristic of autophagy, was detected by AO (Sigma) staining.
Cytometry:Article Title: Penfluridol triggers cytoprotective autophagy and cellular apoptosis through ROS induction and activation of the PP2A-modulated MAPK pathway in acute myeloid leukemia with different FLT3 statuses
Article Snippet: Densitometric analysis of the bands was carried out using Image-Pro Plus software (Media Cybernetics, Rockville, MD). .. Measurement of reactive oxygen species (ROS) production AML cells were treated with penfluridol or H2 O2 for the indicated time points, and the intracellular ROS production was detected by staining with the fluoroprobe, 2′,7′-dichlorofluorescin diacetate (DCFDA, Sigma), in RPMI 1640 medium for 30 min. After washing cells with PBS, ROS production of DCFDA-preloaded cells was captured using a fluorescence microscope (Zeiss Axioplan) or quantified by flow cytometry. .. Detection and quantification of autophagic cells by staining with acridine orange (AO) Formation of acidic vesicular organelles (AVOs), a morphological characteristic of autophagy, was detected by AO (Sigma) staining.
MTT Assay:Article Title: Exendin-4 Pretreated Adipose Derived Stem Cells Are Resistant to Oxidative Stress and Improve Cardiac Performance via Enhanced Adhesion in the Infarcted Heart
Article Snippet: Cell proliferation assay ADSCs were plated in a 96-well plate at a density of 1×104 /well. .. After pretreatment with 50 nM Exendin-4, H2 O2 was given to the cells for 6 h, a total of 5 g/L 3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) (Sigma) in medium was added to the cells for 2 h. Then the medium was removed and dimethyl sulphoxide (DMSO) was applied at a volume of 200 µL/well. .. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) ADSCs that were treated by H2 O2 with or without Exendin-4 were collected.
Activity Assay:Article Title: Hybrid caffeic acid derivatives as monoamine oxidases inhibitors: synthesis, radical scavenging activity, molecular docking studies and in silico ADMET analysis
Article Snippet: Materials and methods Unless otherwise noted, the chemicals required for synthesis and antioxidant activity were purchased from Hi-media Laboratories. .. The biological hMAO activity evaluation of the test drugs was examined by quantifying their action on the generation of H2 O2 by p-tyramine (general substrate for hMAO-B and hMAO-A), utilizing the Amplex Red MAO assay kit (Sigma USA) and MAO isoforms (microsomal) obtained from insect cells (BTI-TN-5B14) expressed as recombinant baculovirus consisting cDNA probes for hMAO-A or hMAO-B. .. Reactions were monitored by thin layer chromatography TLC executed over silica gel precoated plates (0.25 mm) purchased from Merck, envisioned of single spots was carried in iodine and UV chambers, in mobile media TLC- Benzene:Chloroform (7:3).
Recombinant:Article Title: Hybrid caffeic acid derivatives as monoamine oxidases inhibitors: synthesis, radical scavenging activity, molecular docking studies and in silico ADMET analysis
Article Snippet: Materials and methods Unless otherwise noted, the chemicals required for synthesis and antioxidant activity were purchased from Hi-media Laboratories. .. The biological hMAO activity evaluation of the test drugs was examined by quantifying their action on the generation of H2 O2 by p-tyramine (general substrate for hMAO-B and hMAO-A), utilizing the Amplex Red MAO assay kit (Sigma USA) and MAO isoforms (microsomal) obtained from insect cells (BTI-TN-5B14) expressed as recombinant baculovirus consisting cDNA probes for hMAO-A or hMAO-B. .. Reactions were monitored by thin layer chromatography TLC executed over silica gel precoated plates (0.25 mm) purchased from Merck, envisioned of single spots was carried in iodine and UV chambers, in mobile media TLC- Benzene:Chloroform (7:3).
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