96 well flat bottom plates Thermo Fisher Search Results


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  • 99
    Thermo Fisher flat bottom immuno plates
    A standard curve with rHfIL-1β by dehydration coating method. To establish an ELISA system for HfIL-1β, first a standard curve was established with rHfIL-1β using the dehydration coating method. The purified rHfIL-1β was diluted with the coating buffer with the following final concentrations: 0, 3, 6, 9, 12, 15, 15, 18, 21, and 24 ng/mL. Fifty-microliter of serially diluted rHfIL-1β were added to each well of Nunc <t>MaxiSorp®</t> flat-bottom 96-well plate, followed by incubation at 60°C for 2 h. Then, the plate was sequentially incubated with anti-ChIL-1β pAb (1:1000) and goat anti-rabbit antibody (1: 2000). The HRP signal was developed with TMB solution for 30 min. The coating buffer itself was used as negative control. Values represent the mean of three independent experiments. Error bars represent standard error of the mean. The dashed line indicates the threshold line, representing the value of negative control and limitation of the developed ELISA system (OD450 = 0.038)
    Flat Bottom Immuno Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher flat bottom plate
    A standard curve with rHfIL-1β by dehydration coating method. To establish an ELISA system for HfIL-1β, first a standard curve was established with rHfIL-1β using the dehydration coating method. The purified rHfIL-1β was diluted with the coating buffer with the following final concentrations: 0, 3, 6, 9, 12, 15, 15, 18, 21, and 24 ng/mL. Fifty-microliter of serially diluted rHfIL-1β were added to each well of Nunc <t>MaxiSorp®</t> flat-bottom 96-well plate, followed by incubation at 60°C for 2 h. Then, the plate was sequentially incubated with anti-ChIL-1β pAb (1:1000) and goat anti-rabbit antibody (1: 2000). The HRP signal was developed with TMB solution for 30 min. The coating buffer itself was used as negative control. Values represent the mean of three independent experiments. Error bars represent standard error of the mean. The dashed line indicates the threshold line, representing the value of negative control and limitation of the developed ELISA system (OD450 = 0.038)
    Flat Bottom Plate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Price from $9.99 to $1999.99
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    91
    Thermo Fisher flat bottom microtiter plates
    Time course experiment to determine WST-1 reduction in the presence and absence of the electron donors glucose, succinate, and pyruvate (16.6 mM each). Mycobacterium (Myc.) and Sphingomonas (Sph.) cells were grown in <t>microtiter</t> plates with PAHs as sole sources of carbon and energy. The error bars represent ±1 standard deviation. Symbols: ▵, PAH plus e-donors; □, PAH with no e-donors; ○, no PAH but with e-donors.
    Flat Bottom Microtiter Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 180 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher flat bottom nunc plate
    Time course experiment to determine WST-1 reduction in the presence and absence of the electron donors glucose, succinate, and pyruvate (16.6 mM each). Mycobacterium (Myc.) and Sphingomonas (Sph.) cells were grown in <t>microtiter</t> plates with PAHs as sole sources of carbon and energy. The error bars represent ±1 standard deviation. Symbols: ▵, PAH plus e-donors; □, PAH with no e-donors; ○, no PAH but with e-donors.
    Flat Bottom Nunc Plate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher 96 well flat bottom
    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in <t>96-well</t> plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    96 Well Flat Bottom, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher flat bottom 96 well plates
    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in <t>96-well</t> plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    Flat Bottom 96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 565 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flat bottom 96 well plates/product/Thermo Fisher
    Average 99 stars, based on 565 article reviews
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    flat bottom 96 well plates - by Bioz Stars, 2020-08
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    92
    Corning Life Sciences 96 well flat bottom plate
    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in <t>96-well</t> plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    96 Well Flat Bottom Plate, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 92/100, based on 223 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher flat bottomed 96 well microlite plates
    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in <t>96-well</t> plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    Flat Bottomed 96 Well Microlite Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher opaque flat bottom 96 well plates
    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in <t>96-well</t> plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    Opaque Flat Bottom 96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher 96 well flat bottom maxisorp plates
    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in <t>96-well</t> plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    96 Well Flat Bottom Maxisorp Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher flat bottom elisa plates
    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in <t>96-well</t> plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    Flat Bottom Elisa Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher flat bottom nunc 96 well plate
    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in <t>96-well</t> plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    Flat Bottom Nunc 96 Well Plate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flat bottom nunc 96 well plate/product/Thermo Fisher
    Average 93 stars, based on 3 article reviews
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    Image Search Results


    A standard curve with rHfIL-1β by dehydration coating method. To establish an ELISA system for HfIL-1β, first a standard curve was established with rHfIL-1β using the dehydration coating method. The purified rHfIL-1β was diluted with the coating buffer with the following final concentrations: 0, 3, 6, 9, 12, 15, 15, 18, 21, and 24 ng/mL. Fifty-microliter of serially diluted rHfIL-1β were added to each well of Nunc MaxiSorp® flat-bottom 96-well plate, followed by incubation at 60°C for 2 h. Then, the plate was sequentially incubated with anti-ChIL-1β pAb (1:1000) and goat anti-rabbit antibody (1: 2000). The HRP signal was developed with TMB solution for 30 min. The coating buffer itself was used as negative control. Values represent the mean of three independent experiments. Error bars represent standard error of the mean. The dashed line indicates the threshold line, representing the value of negative control and limitation of the developed ELISA system (OD450 = 0.038)

    Journal: BMC Veterinary Research

    Article Title: Development and validation of a house finch interleukin-1β (HfIL-1β) ELISA system

    doi: 10.1186/s12917-017-1199-9

    Figure Lengend Snippet: A standard curve with rHfIL-1β by dehydration coating method. To establish an ELISA system for HfIL-1β, first a standard curve was established with rHfIL-1β using the dehydration coating method. The purified rHfIL-1β was diluted with the coating buffer with the following final concentrations: 0, 3, 6, 9, 12, 15, 15, 18, 21, and 24 ng/mL. Fifty-microliter of serially diluted rHfIL-1β were added to each well of Nunc MaxiSorp® flat-bottom 96-well plate, followed by incubation at 60°C for 2 h. Then, the plate was sequentially incubated with anti-ChIL-1β pAb (1:1000) and goat anti-rabbit antibody (1: 2000). The HRP signal was developed with TMB solution for 30 min. The coating buffer itself was used as negative control. Values represent the mean of three independent experiments. Error bars represent standard error of the mean. The dashed line indicates the threshold line, representing the value of negative control and limitation of the developed ELISA system (OD450 = 0.038)

    Article Snippet: Enzyme-linked Immunosorbent assay (ELISA) To develop the ELISA system for HfIL-1β, we adopted a direct ELISA approach using two different coating methods, carbonate and dehydration, on Nunc MaxiSorp® flat-bottom 96-well plates (Thermo Scientific, IL).

    Techniques: Enzyme-linked Immunosorbent Assay, Purification, Incubation, Negative Control

    Time course experiment to determine WST-1 reduction in the presence and absence of the electron donors glucose, succinate, and pyruvate (16.6 mM each). Mycobacterium (Myc.) and Sphingomonas (Sph.) cells were grown in microtiter plates with PAHs as sole sources of carbon and energy. The error bars represent ±1 standard deviation. Symbols: ▵, PAH plus e-donors; □, PAH with no e-donors; ○, no PAH but with e-donors.

    Journal: Applied and Environmental Microbiology

    Article Title: Detection of Microbial Growth on Polycyclic Aromatic Hydrocarbons in Microtiter Plates by Using the Respiration Indicator WST-1

    doi: 10.1128/AEM.68.6.2683-2689.2002

    Figure Lengend Snippet: Time course experiment to determine WST-1 reduction in the presence and absence of the electron donors glucose, succinate, and pyruvate (16.6 mM each). Mycobacterium (Myc.) and Sphingomonas (Sph.) cells were grown in microtiter plates with PAHs as sole sources of carbon and energy. The error bars represent ±1 standard deviation. Symbols: ▵, PAH plus e-donors; □, PAH with no e-donors; ○, no PAH but with e-donors.

    Article Snippet: Sterile, 96-well, flat-bottom microtiter plates were obtained from Nalge Nunc International, Roskilde, Denmark (product no. 269787).

    Techniques: Standard Deviation

    Effects of various electron donors and azide on the respiratory reduction of WST-1 by mycobacteria (Myc.) and sphingomonads (Sph.) grown for 7 days in microtiter plates with PAHs as sole sources of carbon and energy. The plates were incubated with WST-1 for 90 min. The controls included wells with no PAH but with inoculum and the combined e-donors and wells without bacteria but with PAHs and the combined e-donors. The error bars represent ±1 standard deviation.

    Journal: Applied and Environmental Microbiology

    Article Title: Detection of Microbial Growth on Polycyclic Aromatic Hydrocarbons in Microtiter Plates by Using the Respiration Indicator WST-1

    doi: 10.1128/AEM.68.6.2683-2689.2002

    Figure Lengend Snippet: Effects of various electron donors and azide on the respiratory reduction of WST-1 by mycobacteria (Myc.) and sphingomonads (Sph.) grown for 7 days in microtiter plates with PAHs as sole sources of carbon and energy. The plates were incubated with WST-1 for 90 min. The controls included wells with no PAH but with inoculum and the combined e-donors and wells without bacteria but with PAHs and the combined e-donors. The error bars represent ±1 standard deviation.

    Article Snippet: Sterile, 96-well, flat-bottom microtiter plates were obtained from Nalge Nunc International, Roskilde, Denmark (product no. 269787).

    Techniques: Incubation, Standard Deviation

    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in 96-well plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.

    Journal: Journal of Clinical Investigation

    Article Title: Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators

    doi: 10.1172/JCI25901

    Figure Lengend Snippet: Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in 96-well plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.

    Article Snippet: KerTr cells (human transformed skin keratinocyte, obtained from ATCC) were plated in 96-well flat-bottom plates (3000 cells per well) in 100 μl keratinocyte serum-free medium supplemented with bovine pituitary extract in the absence of EGF (Invitrogen Corp.) and incubated at 37°C for 2 days.

    Techniques: BrdU Incorporation Assay, Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Isolation