The Experts below are selected from a list of 300 Experts worldwide ranked by ideXlab platform
Young-soo Keum - One of the best experts on this subject based on the ideXlab platform.
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sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using dna dye conjugate as a Fluorescent Label
Journal of Environmental Monitoring, 2012Co-Authors: Ziyan Fan, Weilin L Shelver, Young-soo Keum, Lianghong GuoAbstract:Indirect competitive immunoassays were developed on protein microarrays for the sensitive and simultaneous detection of multiple environmental chemicals in one sample. In this assay, a DNA/SYTOX Orange conjugate was employed as an antibody Label to increase the fluorescence signal and sensitivity of the immunoassays. Epoxy-modified glass slides were selected as the substrate for the production of 4 × 4 coating antigen microarrays. With this signal-enhancing system, competition curves for 17β-estradiol (E2), benzo[a]pyrene (BaP) and 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) were obtained individually on the protein microarray. The IC50 and calculated limit of detection (LOD) are 0.32 μg L−1 and 0.022 μg L−1 for E2, 37.2 μg L−1 and 24.5 μg L−1 for BaP, and 31.6 μg L−1 and 2.8 μg L−1 for BDE-47, respectively. LOD of E2 is 14-fold lower than the value reported in a previous study using Cy3 Labeled antibody (Du et al., Clin. Chem, 2005, 51, 368–375). The results of the microarray immunoassay were within 15% of chromatographic analysis for all three pollutants in spiked river water samples, thus verifying the immunoassay. Simultaneous detection of E2, BaP and BDE-47 in one sample was demonstrated. There was no cross-reaction in the immunoassay between these three environmental chemicals. These results suggest that microarray-based immunoassays with DNA/dye conjugate Labels are useful tools for the rapid, sensitive, and high throughput screening of multiple environmental contaminants.
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Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a Fluorescent Label
Journal of Environmental Monitoring, 2012Co-Authors: Young-soo Keum, Qing X. Li, Weilin L ShelverAbstract:Indirect competitive immunoassays were developed on protein microarrays for the sensitive and simultaneous detection of multiple environmental chemicals in one sample. In this assay, a DNA/SYTOX Orange conjugate was employed as an antibody Label to increase the fluorescence signal and sensitivity of the immunoassays. Epoxy-modified glass slides were selected as the substrate for the production of 4 × 4 coating antigen microarrays. With this signal-enhancing system, competition curves for 17β-estradiol (E2), benzo[a]pyrene (BaP) and 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) were obtained individually on the protein microarray. The IC50 and calculated limit of detection (LOD) are 0.32 μg L−1 and 0.022 μg L−1 for E2, 37.2 μg L−1 and 24.5 μg L−1 for BaP, and 31.6 μg L−1 and 2.8 μg L−1 for BDE-47, respectively. LOD of E2 is 14-fold lower than the value reported in a previous study using Cy3 Labeled antibody (Du et al., Clin. Chem, 2005, 51, 368–375). The results of the microarray immunoassay were within 15% of chromatographic analysis for all three pollutants in spiked river water samples, thus verifying the immunoassay. Simultaneous detection of E2, BaP and BDE-47 in one sample was demonstrated. There was no cross-reaction in the immunoassay between these three environmental chemicals. These results suggest that microarray-based immunoassays with DNA/dye conjugate Labels are useful tools for the rapid, sensitive, and high throughput screening of multiple environmental contaminants.
V Metelskaya - One of the best experts on this subject based on the ideXlab platform.
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direct Labeling of serum proteins by Fluorescent dye for antibody microarray
Biochemical and Biophysical Research Communications, 2017Co-Authors: M V Klimushina, Nadezhda G Gumanova, V MetelskayaAbstract:Abstract Analysis of serum proteome by antibody microarray is used to identify novel biomarkers and to study signaling pathways including protein phosphorylation and protein-protein interactions. Labeling of serum proteins is important for optimal performance of the antibody microarray. Proper choice of Fluorescent Label and optimal concentration of protein loaded on the microarray ensure good quality of imaging that can be reliably scanned and processed by the software. We have optimized direct serum protein Labeling using Fluorescent dye Arrayit Green 540 (Arrayit Corporation, USA) for antibody microarray. Optimized procedure produces high quality images that can be readily scanned and used for statistical analysis of protein composition of the serum.
Marco Fritzsche - One of the best experts on this subject based on the ideXlab platform.
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statistical analysis of scanning fluorescence correlation spectroscopy data differentiates free from hindered diffusion
ACS Nano, 2018Co-Authors: Falk Schneider, Dominic Waithe, B C Lagerholm, Dilip Shrestha, Erdinc Sezgin, Christian Eggeling, Marco FritzscheAbstract:Cells rely on versatile diffusion dynamics in their plasma membrane. Quantification of this often heterogeneous diffusion is essential to the understanding of cell regulation and function. Yet such measurements remain a major challenge in cell biology, usually due to low sampling throughput, a necessity for dedicated equipment, sophisticated Fluorescent Label strategies, and limited sensitivity. Here, we introduce a robust, broadly applicable statistical analysis pipeline for large scanning fluorescence correlation spectroscopy data sets, which uncovers the nanoscale heterogeneity of the plasma membrane in living cells by differentiating free from hindered diffusion modes of Fluorescent lipid and protein analogues.
Weilin L Shelver - One of the best experts on this subject based on the ideXlab platform.
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sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using dna dye conjugate as a Fluorescent Label
Journal of Environmental Monitoring, 2012Co-Authors: Ziyan Fan, Weilin L Shelver, Young-soo Keum, Lianghong GuoAbstract:Indirect competitive immunoassays were developed on protein microarrays for the sensitive and simultaneous detection of multiple environmental chemicals in one sample. In this assay, a DNA/SYTOX Orange conjugate was employed as an antibody Label to increase the fluorescence signal and sensitivity of the immunoassays. Epoxy-modified glass slides were selected as the substrate for the production of 4 × 4 coating antigen microarrays. With this signal-enhancing system, competition curves for 17β-estradiol (E2), benzo[a]pyrene (BaP) and 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) were obtained individually on the protein microarray. The IC50 and calculated limit of detection (LOD) are 0.32 μg L−1 and 0.022 μg L−1 for E2, 37.2 μg L−1 and 24.5 μg L−1 for BaP, and 31.6 μg L−1 and 2.8 μg L−1 for BDE-47, respectively. LOD of E2 is 14-fold lower than the value reported in a previous study using Cy3 Labeled antibody (Du et al., Clin. Chem, 2005, 51, 368–375). The results of the microarray immunoassay were within 15% of chromatographic analysis for all three pollutants in spiked river water samples, thus verifying the immunoassay. Simultaneous detection of E2, BaP and BDE-47 in one sample was demonstrated. There was no cross-reaction in the immunoassay between these three environmental chemicals. These results suggest that microarray-based immunoassays with DNA/dye conjugate Labels are useful tools for the rapid, sensitive, and high throughput screening of multiple environmental contaminants.
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Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a Fluorescent Label
Journal of Environmental Monitoring, 2012Co-Authors: Young-soo Keum, Qing X. Li, Weilin L ShelverAbstract:Indirect competitive immunoassays were developed on protein microarrays for the sensitive and simultaneous detection of multiple environmental chemicals in one sample. In this assay, a DNA/SYTOX Orange conjugate was employed as an antibody Label to increase the fluorescence signal and sensitivity of the immunoassays. Epoxy-modified glass slides were selected as the substrate for the production of 4 × 4 coating antigen microarrays. With this signal-enhancing system, competition curves for 17β-estradiol (E2), benzo[a]pyrene (BaP) and 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) were obtained individually on the protein microarray. The IC50 and calculated limit of detection (LOD) are 0.32 μg L−1 and 0.022 μg L−1 for E2, 37.2 μg L−1 and 24.5 μg L−1 for BaP, and 31.6 μg L−1 and 2.8 μg L−1 for BDE-47, respectively. LOD of E2 is 14-fold lower than the value reported in a previous study using Cy3 Labeled antibody (Du et al., Clin. Chem, 2005, 51, 368–375). The results of the microarray immunoassay were within 15% of chromatographic analysis for all three pollutants in spiked river water samples, thus verifying the immunoassay. Simultaneous detection of E2, BaP and BDE-47 in one sample was demonstrated. There was no cross-reaction in the immunoassay between these three environmental chemicals. These results suggest that microarray-based immunoassays with DNA/dye conjugate Labels are useful tools for the rapid, sensitive, and high throughput screening of multiple environmental contaminants.
Ursula Sauer - One of the best experts on this subject based on the ideXlab platform.
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Quality control of chip manufacture and chip analysis using epoxy-chips as a model
Sensors and Actuators B-chemical, 2003Co-Authors: Claudia Preininger, Ursula SauerAbstract:Abstract Biochips are miniaturized, highly ordered analysis systems which offer the unique advantage of highly parallel analysis of thousands of analytes at the same time. Although this technique has been enthusiastically developed and has promised to improve and speed up numerous biological assays, the quality control of chip manufacture, chip analysis and data management has received less attention. The following article compares three epoxy-containing chip surfaces (ARChip Epoxy, 3D-Link™, and EasySpot) with respect to their autofluorescence, immobilization capacity, background fluorescence and hybridization efficiency. Since data collected from biochip experiments are random snapshots with errors, inherently noisy and incomplete, we tried to evaluate technical factors causing variability and to set up quality control procedures for chip manufacture and chip analysis. Variabilities caused by arraying, glass substrate and polymer coating, Fluorescent Label and experimental conditions are discussed in details.