96 well flat bottom plates Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher 96 well flat bottom plates
    Effect of TPT on the proliferative response of bidirectional MLR and anti-CD3/CD28–stimulated CD4 + T cells. Allogeneic human PBMCs from 2 different donors (3 × 10 5 cells/well each) were cocultured in <t>96-well</t> flat-bottom plates in the presence
    96 Well Flat Bottom Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 771 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/96 well flat bottom plates/product/Thermo Fisher
    Average 99 stars, based on 771 article reviews
    Price from $9.99 to $1999.99
    96 well flat bottom plates - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Millipore 96 well flat bottom plates
    IFN- γ Elispot response to mitogens in lean, overweight, and obese chimpanzees. PBMCs isolated from the blood samples of the chimpanzees. Triplicate wells of the <t>96-well</t> microtiter plates, precoated with IFN- γ antibody, were seeded with 10 5 PBMCs from the monkeys and incubated with 5 μ g of each of the mitogens for 36 h at 37°C. At the end of the incubation period, the wells were washed and stained with biotinylated second IFN- γ antibody. The total number of spot forming cells in each of the mitogen-stimulated wells was counted and adjusted to control medium as background. See method section for experiment details. The results shown are an average of two distinct experiments and comparison between groups of chimpanzees was done by one-way analysis of variance with the Kruskal-Wallis test as described in Figure 1 .
    96 Well Flat Bottom Plates, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 522 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/96 well flat bottom plates/product/Millipore
    Average 99 stars, based on 522 article reviews
    Price from $9.99 to $1999.99
    96 well flat bottom plates - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    91
    Thermo Fisher clear flat bottom immuno nonsterile 96 well plates
    IFN- γ Elispot response to mitogens in lean, overweight, and obese chimpanzees. PBMCs isolated from the blood samples of the chimpanzees. Triplicate wells of the <t>96-well</t> microtiter plates, precoated with IFN- γ antibody, were seeded with 10 5 PBMCs from the monkeys and incubated with 5 μ g of each of the mitogens for 36 h at 37°C. At the end of the incubation period, the wells were washed and stained with biotinylated second IFN- γ antibody. The total number of spot forming cells in each of the mitogen-stimulated wells was counted and adjusted to control medium as background. See method section for experiment details. The results shown are an average of two distinct experiments and comparison between groups of chimpanzees was done by one-way analysis of variance with the Kruskal-Wallis test as described in Figure 1 .
    Clear Flat Bottom Immuno Nonsterile 96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 505 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/clear flat bottom immuno nonsterile 96 well plates/product/Thermo Fisher
    Average 91 stars, based on 505 article reviews
    Price from $9.99 to $1999.99
    clear flat bottom immuno nonsterile 96 well plates - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    92
    Corning Life Sciences 96 well flat bottom plates
    Configuration of the <t>96-well</t> flat bottom plates in which the cytotoxicity assay was carried out. M = complete medium without cells. A = complete medium without extract solutions. Sa = artificial saliva (negative control). T = TNF (positive control). C1, C2, and C3 = mini-implants Conexão, tested in triplicate. N1, N2, and N3 = mini-implants Neodent, tested in triplicate. S1, S2, and S3 = mini-implants SIN, tested in triplicate. The yellow wells remained empty.
    96 Well Flat Bottom Plates, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 92/100, based on 669 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/96 well flat bottom plates/product/Corning Life Sciences
    Average 92 stars, based on 669 article reviews
    Price from $9.99 to $1999.99
    96 well flat bottom plates - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    92
    Becton Dickinson 96 well flat bottom plates
    Assessment of cell death, apoptosis and cell cycle in SKBR3, JIMT-1 and MCF7-HER2 cells treated with gefitinib, RAD001 and the gefitinib and RAD001 combination used at 200:1 (Gef:RAD) molar ratio . (A) Assessment of cell death by HCS. Cells were seeded in <t>96-well</t> plates and treated with indicated drug concentration. After 72 h cells were stained in situ with DRAQ5 (stain for viable cells) and ethidium homodimer (stain for dead cells) and images were acquired with IN Cell 1000. The imaging data were analyzed with the IN Cell 1000 Investigator software and the results are expressed as percentage of dead cells relative to DMSO control. Asterisks above data points indicate a significantly (p
    96 Well Flat Bottom Plates, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 723 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/96 well flat bottom plates/product/Becton Dickinson
    Average 92 stars, based on 723 article reviews
    Price from $9.99 to $1999.99
    96 well flat bottom plates - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    92
    Corning Life Sciences flat bottom 96 well plates
    Synergistic activity of anti-folate combinations against E . coli and S . aureus . a Targets of anti-folate compounds. b – d E . coli BW25113 strain was grown overnight in LB medium. Cultures were washed twice and resuspended in M9-glucose, then inoculated into a <t>96-well</t> round-bottom plate (Corning) containing the same medium with a range of concentrations of SMX, TMP, MAC173979, or combination of two compounds. Concentration ranges were as follows: SMX (0.024–25 µg ml −1 ), TMP (0.0078–1 µg ml −1 ), and MAC173979 (0.05–25 µg ml −1 ). MICs were determined by visible growth after 24 h incubation at 37 °C. Synergy was assessed by calculating FICI. FICI m , minimum value of FICI in the tested combinations is shown. Synergy (FICI m ≤ 0.5). No interaction (FICI m > 0.5). b – d Graphical representations of E . coli BW25113 checkerboard assays are shown. Representative data from at least three independent experiments are shown. b SMX and MAC173979. c TMP and MAC173979. d SMX and TMP
    Flat Bottom 96 Well Plates, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 92/100, based on 457 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flat bottom 96 well plates/product/Corning Life Sciences
    Average 92 stars, based on 457 article reviews
    Price from $9.99 to $1999.99
    flat bottom 96 well plates - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    99
    Thermo Fisher flat bottom 96 well plates
    Synergistic activity of anti-folate combinations against E . coli and S . aureus . a Targets of anti-folate compounds. b – d E . coli BW25113 strain was grown overnight in LB medium. Cultures were washed twice and resuspended in M9-glucose, then inoculated into a <t>96-well</t> round-bottom plate (Corning) containing the same medium with a range of concentrations of SMX, TMP, MAC173979, or combination of two compounds. Concentration ranges were as follows: SMX (0.024–25 µg ml −1 ), TMP (0.0078–1 µg ml −1 ), and MAC173979 (0.05–25 µg ml −1 ). MICs were determined by visible growth after 24 h incubation at 37 °C. Synergy was assessed by calculating FICI. FICI m , minimum value of FICI in the tested combinations is shown. Synergy (FICI m ≤ 0.5). No interaction (FICI m > 0.5). b – d Graphical representations of E . coli BW25113 checkerboard assays are shown. Representative data from at least three independent experiments are shown. b SMX and MAC173979. c TMP and MAC173979. d SMX and TMP
    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
    Price from $9.99 to $1999.99
    flat bottom 96 well plates - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Becton Dickinson 96 well flat bottom plate
    CXCR7 regulates multiple functions in HBMECs. A.  shRNA-scramble CXCR7 or shRNA-CXCR7 HBMECs were seeded on 96 well plate in the presence of 200 ng/ml CXCL12 and cell proliferation was analyzed by XTT assay as described in “  Materials and Methods ”.  B.  shRNA-scramble or shRNA-CXCR7 HBMECs were seeded on 24 well plate coated with matrigel and tubulogenesis assay was performed as described in in “  Materials and Methods ”.  C.  shRNA-scramble CXCR7 or shRNA-CXCR7 HBMECs were seeded on the upper chamber of BD matrigel invasion chambers while the lower chamber was filled with 600 µl 10% complete medium with CXCL12 (200 ng/ml). Cell migration ability was analyzed as described in “  Materials and Methods ”. Data are shown as means ± S.D from three independent experiments. *,  p
    96 Well Flat Bottom Plate, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 414 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/96 well flat bottom plate/product/Becton Dickinson
    Average 93 stars, based on 414 article reviews
    Price from $9.99 to $1999.99
    96 well flat bottom plate - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    93
    Greiner Bio flat bottom 96 well plates
    Isolation of genotypically and phenotypically diverse single stem-like cell-derived subclones of paediatric GBM and DIPG. (A) Isolation of subclonal populations. Experimental schema for disaggregation of heterogeneous mixtures of patient-derived tumour cells, flow sorting into single cells in <t>96-well</t> plates, and allowing colonies to form as either 2D cultures adherent on laminin, or 3D neurospheres, all under stem cell conditions. Individual subclonal colonies are subjected to high-throughput phenotypic analysis and targeted resequencing, and further cultured for detailed in vitro and in vivo mechanistic comparison with heterogeneous bulk populations. (B) Clonogenicity. Percentage of single cells which formed colonies under 2D laminin and 3D neurosphere stem cell conditions are given for six pGBM and DIPG primary patient-derived cell cultures, labelled by anatomical location and histone H3 mutation subgroup. (C) 3D neurosphere culture from single cell-derived colonies from SU-DIPG-VI assessed by Celigo S imaging cytometer. (D) Growth of single cell-derived colonies over time, assessed as diammeter of neurosphere, labelled and colour-coded. (E) Targeted sequencing contingency plot of somatic mutations identified as common to all subclones (blue), shared amongst certain subclones (yellow) and private to individuals (red). (F) 2D laminin culture from single cell-derived colonies from SU-DIPG-VI assessed by Celigo S imaging cytometer. (G) Growth of single cell-derived colonies over time, assessed as diammeter of neurosphere, with subclones taken for later analysis highlighted: A-D10 (fast, purple), A-B8 (intermediate, pink) and A-E6 (slow, violet). (H) Targeted sequencing contingency plot of somatic mutations identified as common to all subclones (blue), shared amongst certain subclones (yellow) and private to individuals (red). Gene names are coloured to highlight private mutations in selected subclones, or common to A-D10 and A-B8 (brown). (I) Growth. Time-course for growth of selected subclones re-plated and grown over 160 hours, highlighting statistically significant differences among subclones and heterogeneous bulk cell populations of SU-DIPG-VI (blue). Representative images at 72 hours are provided from the Celigo S cytometer, with tumour cells marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. (J) Invasion. Time-course of invasion of cells into a matrigel matrix over 72 hours, either as percentage of the total area in the field of view covered by invading cells, or as a percentage of time zero. Representative images given at 72 hours, with extent of tumour cell invasion marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. (K) Migration. Time-course of tumour cell migration onto matrigel over 72 hours, either as percentage of the total area of the well covered by migrating cells, or as a percentage of time zero. Representative images given at 72 hours, with extent of tumour cell migration marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. * p
    Flat Bottom 96 Well Plates, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 93/100, based on 167 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flat bottom 96 well plates/product/Greiner Bio
    Average 93 stars, based on 167 article reviews
    Price from $9.99 to $1999.99
    flat bottom 96 well plates - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    91
    Corning Life Sciences 96 well flat bottom microtiter plates
    In vitro characterization of the inhibitory activity of the leading compound 9029936 on C. albicans biofilm formation. (A) Dose-dependent inhibitory effects of compound 9029936 on C. albicans biofilm formation. The compound was tested in serial 2-fold dilutions (concentrations ranging from 40 to 0.078 µM), with appropriate positive and negative controls. Results shown are the mean percent biofilm inhibition relative to control biofilms (grown in the absence of compound 9029936), determined in XTT colorimetric assays for multiple technical replicates from several independent experiments. Error bars indicate standard deviations. (B) The leading compound inhibited the proliferation and maturation phases of C. albicans biofilm development. A biofilm kinetic assay was performed in order to examine the inhibitory effects of compound 9029936 at different stages of biofilm development, using the same <t>96-well</t> <t>microtiter</t> plate model of C. albicans biofilm formation. The extent of inhibition was determined at multiple times (2, 6, 8, 12, 24, and 48 h) after seeding the wells with C. albicans cells in the presence or absence of compound 9029936 (5 μM concentration). Results are expressed as means of multiple technical replicates from a single experiment, with error bars representing standard deviations. (C) SEM images of biofilms of C. albicans strain SC5314 formed in the absence or presence of compound 9029936 at a concentration of 5 μM. Bars, 20 μm.
    96 Well Flat Bottom Microtiter Plates, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 91/100, based on 100 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/96 well flat bottom microtiter plates/product/Corning Life Sciences
    Average 91 stars, based on 100 article reviews
    Price from $9.99 to $1999.99
    96 well flat bottom microtiter plates - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    84
    Thermo Fisher procartaplex 96 well flat bottom plate
    In vitro characterization of the inhibitory activity of the leading compound 9029936 on C. albicans biofilm formation. (A) Dose-dependent inhibitory effects of compound 9029936 on C. albicans biofilm formation. The compound was tested in serial 2-fold dilutions (concentrations ranging from 40 to 0.078 µM), with appropriate positive and negative controls. Results shown are the mean percent biofilm inhibition relative to control biofilms (grown in the absence of compound 9029936), determined in XTT colorimetric assays for multiple technical replicates from several independent experiments. Error bars indicate standard deviations. (B) The leading compound inhibited the proliferation and maturation phases of C. albicans biofilm development. A biofilm kinetic assay was performed in order to examine the inhibitory effects of compound 9029936 at different stages of biofilm development, using the same <t>96-well</t> <t>microtiter</t> plate model of C. albicans biofilm formation. The extent of inhibition was determined at multiple times (2, 6, 8, 12, 24, and 48 h) after seeding the wells with C. albicans cells in the presence or absence of compound 9029936 (5 μM concentration). Results are expressed as means of multiple technical replicates from a single experiment, with error bars representing standard deviations. (C) SEM images of biofilms of C. albicans strain SC5314 formed in the absence or presence of compound 9029936 at a concentration of 5 μM. Bars, 20 μm.
    Procartaplex 96 Well Flat Bottom Plate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/procartaplex 96 well flat bottom plate/product/Thermo Fisher
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    procartaplex 96 well flat bottom plate - by Bioz Stars, 2020-09
    84/100 stars
      Buy from Supplier

    99
    Thermo Fisher nunc maxisorp flat bottom 96 well 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)
    Nunc Maxisorp Flat Bottom 96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nunc maxisorp flat bottom 96 well plates/product/Thermo Fisher
    Average 99 stars, based on 63 article reviews
    Price from $9.99 to $1999.99
    nunc maxisorp flat bottom 96 well plates - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Effect of TPT on the proliferative response of bidirectional MLR and anti-CD3/CD28–stimulated CD4 + T cells. Allogeneic human PBMCs from 2 different donors (3 × 10 5 cells/well each) were cocultured in 96-well flat-bottom plates in the presence

    Journal:

    Article Title: Triptolide, a constituent of immunosuppressive Chinese herbal medicine, is a potent suppressor of dendritic-cell maturation and trafficking

    doi: 10.1182/blood-2005-03-0854

    Figure Lengend Snippet: Effect of TPT on the proliferative response of bidirectional MLR and anti-CD3/CD28–stimulated CD4 + T cells. Allogeneic human PBMCs from 2 different donors (3 × 10 5 cells/well each) were cocultured in 96-well flat-bottom plates in the presence

    Article Snippet: Purified CD4+ T cells (105 /well) were cultured in 96-well flat-bottom plates with CD3/CD28 T-cell expansion Dynabeads (5 × 103 /well; Dynal ASA, Oslo, Norway).

    Techniques:

    Bacterial survival in mouse blood and biofilm formation ability . (a) The ccs4 gene deletion had no effects on pneumococcal survival in mouse blood. Bacteria were incubated in heparinized mouse blood at 37°C for 1, 2, or 3 hours in a 5% CO 2 atmosphere. Survival rate was calculated by dividing the CFU value after the period of incubation by the CFU value of the original inoculum. Values are presented as the mean of 6 wells from one of 3 independent experiments. Vertical lines represent the mean + S.E. (b) Effect of ccs4 deletion on pneumococcal biofilm formation was assessed following incubation in THY at 37°C for 16 hours. Biofilm formed in 96-well microtiter plates was stained with Gram’s stain solution and absorbance was measured at 550 nm. The experiments were performed 3 times and data shown represent the mean of 6 wells from one of 3 independent experiments. Data are displayed as a box-whisker plot (min-[lower quartile–median-upper quartile]-max). Statistical differences between groups were analyzed using Mann-Whitney’s U -test. (c) Representative microscopic images of bacterial aggregation from WT strain-derived biofilm (left), and Δ ccs4 strain mutant-derived biofilm (right). Scale bars indicate 100 μm.

    Journal: Virulence

    Article Title: Competence-induced protein Ccs4 facilitates pneumococcal invasion into brain tissue and virulence in meningitis

    doi: 10.1080/21505594.2018.1526530

    Figure Lengend Snippet: Bacterial survival in mouse blood and biofilm formation ability . (a) The ccs4 gene deletion had no effects on pneumococcal survival in mouse blood. Bacteria were incubated in heparinized mouse blood at 37°C for 1, 2, or 3 hours in a 5% CO 2 atmosphere. Survival rate was calculated by dividing the CFU value after the period of incubation by the CFU value of the original inoculum. Values are presented as the mean of 6 wells from one of 3 independent experiments. Vertical lines represent the mean + S.E. (b) Effect of ccs4 deletion on pneumococcal biofilm formation was assessed following incubation in THY at 37°C for 16 hours. Biofilm formed in 96-well microtiter plates was stained with Gram’s stain solution and absorbance was measured at 550 nm. The experiments were performed 3 times and data shown represent the mean of 6 wells from one of 3 independent experiments. Data are displayed as a box-whisker plot (min-[lower quartile–median-upper quartile]-max). Statistical differences between groups were analyzed using Mann-Whitney’s U -test. (c) Representative microscopic images of bacterial aggregation from WT strain-derived biofilm (left), and Δ ccs4 strain mutant-derived biofilm (right). Scale bars indicate 100 μm.

    Article Snippet: Overnight bacterial cultures (20 μL) were mixed with 180 μL of THY in 96-well (flat-bottom) microtiter plates (Thermo Fisher Scientific).

    Techniques: Incubation, Staining, Whisker Assay, MANN-WHITNEY, Derivative Assay, Mutagenesis

    IFN- γ Elispot response to mitogens in lean, overweight, and obese chimpanzees. PBMCs isolated from the blood samples of the chimpanzees. Triplicate wells of the 96-well microtiter plates, precoated with IFN- γ antibody, were seeded with 10 5 PBMCs from the monkeys and incubated with 5 μ g of each of the mitogens for 36 h at 37°C. At the end of the incubation period, the wells were washed and stained with biotinylated second IFN- γ antibody. The total number of spot forming cells in each of the mitogen-stimulated wells was counted and adjusted to control medium as background. See method section for experiment details. The results shown are an average of two distinct experiments and comparison between groups of chimpanzees was done by one-way analysis of variance with the Kruskal-Wallis test as described in Figure 1 .

    Journal: International Journal of Inflammation

    Article Title: Obesity Related Alterations in Plasma Cytokines and Metabolic Hormones in Chimpanzees

    doi: 10.1155/2014/856749

    Figure Lengend Snippet: IFN- γ Elispot response to mitogens in lean, overweight, and obese chimpanzees. PBMCs isolated from the blood samples of the chimpanzees. Triplicate wells of the 96-well microtiter plates, precoated with IFN- γ antibody, were seeded with 10 5 PBMCs from the monkeys and incubated with 5 μ g of each of the mitogens for 36 h at 37°C. At the end of the incubation period, the wells were washed and stained with biotinylated second IFN- γ antibody. The total number of spot forming cells in each of the mitogen-stimulated wells was counted and adjusted to control medium as background. See method section for experiment details. The results shown are an average of two distinct experiments and comparison between groups of chimpanzees was done by one-way analysis of variance with the Kruskal-Wallis test as described in Figure 1 .

    Article Snippet: All assays were conducted according to the manufacturer's instructions using handheld magnetic separator block for 96-well flat bottom plates (Millipore, Millipore Corp.) and analyzed using the Luminex 200 system (Bio-Rad Corp.).

    Techniques: Enzyme-linked Immunospot, Isolation, Incubation, Staining

    Configuration of the 96-well flat bottom plates in which the cytotoxicity assay was carried out. M = complete medium without cells. A = complete medium without extract solutions. Sa = artificial saliva (negative control). T = TNF (positive control). C1, C2, and C3 = mini-implants Conexão, tested in triplicate. N1, N2, and N3 = mini-implants Neodent, tested in triplicate. S1, S2, and S3 = mini-implants SIN, tested in triplicate. The yellow wells remained empty.

    Journal: Dental Press Journal of Orthodontics

    Article Title: Evaluation of cytotoxicity and corrosion resistance of orthodontic mini-implants

    doi: 10.1590/2177-6709.21.5.039-046.oar

    Figure Lengend Snippet: Configuration of the 96-well flat bottom plates in which the cytotoxicity assay was carried out. M = complete medium without cells. A = complete medium without extract solutions. Sa = artificial saliva (negative control). T = TNF (positive control). C1, C2, and C3 = mini-implants Conexão, tested in triplicate. N1, N2, and N3 = mini-implants Neodent, tested in triplicate. S1, S2, and S3 = mini-implants SIN, tested in triplicate. The yellow wells remained empty.

    Article Snippet: Cytotoxicity assays Aliquots of 100 µL of L929 cell suspension were pipetted into 96-well flat bottom plates (Corning Costar Corporation, Cambridge, MA, USA).

    Techniques: Cytotoxicity Assay, Negative Control, Positive Control

    Assessment of cell death, apoptosis and cell cycle in SKBR3, JIMT-1 and MCF7-HER2 cells treated with gefitinib, RAD001 and the gefitinib and RAD001 combination used at 200:1 (Gef:RAD) molar ratio . (A) Assessment of cell death by HCS. Cells were seeded in 96-well plates and treated with indicated drug concentration. After 72 h cells were stained in situ with DRAQ5 (stain for viable cells) and ethidium homodimer (stain for dead cells) and images were acquired with IN Cell 1000. The imaging data were analyzed with the IN Cell 1000 Investigator software and the results are expressed as percentage of dead cells relative to DMSO control. Asterisks above data points indicate a significantly (p

    Journal: BMC Cancer

    Article Title: The combination of gefitinib and RAD001 inhibits growth of HER2 overexpressing breast cancer cells and tumors irrespective of trastuzumab sensitivity

    doi: 10.1186/1471-2407-11-420

    Figure Lengend Snippet: Assessment of cell death, apoptosis and cell cycle in SKBR3, JIMT-1 and MCF7-HER2 cells treated with gefitinib, RAD001 and the gefitinib and RAD001 combination used at 200:1 (Gef:RAD) molar ratio . (A) Assessment of cell death by HCS. Cells were seeded in 96-well plates and treated with indicated drug concentration. After 72 h cells were stained in situ with DRAQ5 (stain for viable cells) and ethidium homodimer (stain for dead cells) and images were acquired with IN Cell 1000. The imaging data were analyzed with the IN Cell 1000 Investigator software and the results are expressed as percentage of dead cells relative to DMSO control. Asterisks above data points indicate a significantly (p

    Article Snippet: Alamar Blue and IN Cell 1000 screening assays Cells were plated under standard serum conditions (10% FBS) in their respective media in triplicate wells/condition in 96-well flat bottom plates (Optilux, Falcon, Becton-Dickinson).

    Techniques: Concentration Assay, Staining, In Situ, Imaging, Software

    Synergistic activity of anti-folate combinations against E . coli and S . aureus . a Targets of anti-folate compounds. b – d E . coli BW25113 strain was grown overnight in LB medium. Cultures were washed twice and resuspended in M9-glucose, then inoculated into a 96-well round-bottom plate (Corning) containing the same medium with a range of concentrations of SMX, TMP, MAC173979, or combination of two compounds. Concentration ranges were as follows: SMX (0.024–25 µg ml −1 ), TMP (0.0078–1 µg ml −1 ), and MAC173979 (0.05–25 µg ml −1 ). MICs were determined by visible growth after 24 h incubation at 37 °C. Synergy was assessed by calculating FICI. FICI m , minimum value of FICI in the tested combinations is shown. Synergy (FICI m ≤ 0.5). No interaction (FICI m > 0.5). b – d Graphical representations of E . coli BW25113 checkerboard assays are shown. Representative data from at least three independent experiments are shown. b SMX and MAC173979. c TMP and MAC173979. d SMX and TMP

    Journal: Nature Communications

    Article Title: Mutual potentiation drives synergy between trimethoprim and sulfamethoxazole

    doi: 10.1038/s41467-018-03447-x

    Figure Lengend Snippet: Synergistic activity of anti-folate combinations against E . coli and S . aureus . a Targets of anti-folate compounds. b – d E . coli BW25113 strain was grown overnight in LB medium. Cultures were washed twice and resuspended in M9-glucose, then inoculated into a 96-well round-bottom plate (Corning) containing the same medium with a range of concentrations of SMX, TMP, MAC173979, or combination of two compounds. Concentration ranges were as follows: SMX (0.024–25 µg ml −1 ), TMP (0.0078–1 µg ml −1 ), and MAC173979 (0.05–25 µg ml −1 ). MICs were determined by visible growth after 24 h incubation at 37 °C. Synergy was assessed by calculating FICI. FICI m , minimum value of FICI in the tested combinations is shown. Synergy (FICI m ≤ 0.5). No interaction (FICI m > 0.5). b – d Graphical representations of E . coli BW25113 checkerboard assays are shown. Representative data from at least three independent experiments are shown. b SMX and MAC173979. c TMP and MAC173979. d SMX and TMP

    Article Snippet: The cultures were washed twice and resuspended in M9-glucose then inoculated into each well of a 96-well flat-bottom plate (Corning) containing M9-glucose containing different concentrations of SMX, TMP, or combination of SMX and TMP.

    Techniques: Activity Assay, Concentration Assay, Incubation

    Lipiodol transfected DC are immune modulatory.  A . Day 7 bone marrow derived DC were cultured alone or transfected with mismatched siRNA, or IL-12p35-specific siRNA using 3 μl/culture lipiodol. Following a 24 hour incubation cells were plated at 1 × 10 6  in 6 well culture dishes and activated for 24 hours with 10 ng/ml LPS and 10 ng/ml TNF- α . Supernatant was harvested and analyzed by ELISA for IL-10 production.  B . C57/BL6 DC were transfected with mismatched siRNA, IL-12p35-specific siRNA or lipiodol alone, irradiated (3,000 rad) and seeded in triplicate at various concentrations in a flat-bottom 96-well plate. Splenic T cells from BALB/c mice were added as responders (5 × 10 5  cells/well). The mixed lymphocytes were cultured for 72 h and proliferation was assessed by thymidine incorporation.  C   D . IFN-γ and IL-4 concentrations, respectively, were assessed from MLR cultures at 48 hours of incubation.

    Journal: Journal of Translational Medicine

    Article Title: A novel method of modifying immune responses by vaccination with lipiodol-siRNA mixtures

    doi: 10.1186/1479-5876-4-2

    Figure Lengend Snippet: Lipiodol transfected DC are immune modulatory. A . Day 7 bone marrow derived DC were cultured alone or transfected with mismatched siRNA, or IL-12p35-specific siRNA using 3 μl/culture lipiodol. Following a 24 hour incubation cells were plated at 1 × 10 6 in 6 well culture dishes and activated for 24 hours with 10 ng/ml LPS and 10 ng/ml TNF- α . Supernatant was harvested and analyzed by ELISA for IL-10 production. B . C57/BL6 DC were transfected with mismatched siRNA, IL-12p35-specific siRNA or lipiodol alone, irradiated (3,000 rad) and seeded in triplicate at various concentrations in a flat-bottom 96-well plate. Splenic T cells from BALB/c mice were added as responders (5 × 10 5 cells/well). The mixed lymphocytes were cultured for 72 h and proliferation was assessed by thymidine incorporation. C D . IFN-γ and IL-4 concentrations, respectively, were assessed from MLR cultures at 48 hours of incubation.

    Article Snippet: Mixed lymphocyte reaction C57/BL6 DC after transfection were irradiated (3,000 rad) and seeded in triplicate at various concentrations in a flat-bottom 96-well plate (Corning) for use as stimulator cells.

    Techniques: Transfection, Derivative Assay, Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Irradiation, Mouse Assay

    CXCR7 regulates multiple functions in HBMECs. A.  shRNA-scramble CXCR7 or shRNA-CXCR7 HBMECs were seeded on 96 well plate in the presence of 200 ng/ml CXCL12 and cell proliferation was analyzed by XTT assay as described in “  Materials and Methods ”.  B.  shRNA-scramble or shRNA-CXCR7 HBMECs were seeded on 24 well plate coated with matrigel and tubulogenesis assay was performed as described in in “  Materials and Methods ”.  C.  shRNA-scramble CXCR7 or shRNA-CXCR7 HBMECs were seeded on the upper chamber of BD matrigel invasion chambers while the lower chamber was filled with 600 µl 10% complete medium with CXCL12 (200 ng/ml). Cell migration ability was analyzed as described in “  Materials and Methods ”. Data are shown as means ± S.D from three independent experiments. *,  p

    Journal: PLoS ONE

    Article Title: Chemokine Receptor CXCR7 Is a Functional Receptor for CXCL12 in Brain Endothelial Cells

    doi: 10.1371/journal.pone.0103938

    Figure Lengend Snippet: CXCR7 regulates multiple functions in HBMECs. A. shRNA-scramble CXCR7 or shRNA-CXCR7 HBMECs were seeded on 96 well plate in the presence of 200 ng/ml CXCL12 and cell proliferation was analyzed by XTT assay as described in “ Materials and Methods ”. B. shRNA-scramble or shRNA-CXCR7 HBMECs were seeded on 24 well plate coated with matrigel and tubulogenesis assay was performed as described in in “ Materials and Methods ”. C. shRNA-scramble CXCR7 or shRNA-CXCR7 HBMECs were seeded on the upper chamber of BD matrigel invasion chambers while the lower chamber was filled with 600 µl 10% complete medium with CXCL12 (200 ng/ml). Cell migration ability was analyzed as described in “ Materials and Methods ”. Data are shown as means ± S.D from three independent experiments. *, p

    Article Snippet: Briefly, shRNA-CXCR7 or scramble HBMECs were seeded at 1000 cells/well in 96-well flat bottom plate (BD Bioscience) in the presence of 200 ng/ml CXCL12.

    Techniques: shRNA, XTT Assay, Migration

    Isolation of genotypically and phenotypically diverse single stem-like cell-derived subclones of paediatric GBM and DIPG. (A) Isolation of subclonal populations. Experimental schema for disaggregation of heterogeneous mixtures of patient-derived tumour cells, flow sorting into single cells in 96-well plates, and allowing colonies to form as either 2D cultures adherent on laminin, or 3D neurospheres, all under stem cell conditions. Individual subclonal colonies are subjected to high-throughput phenotypic analysis and targeted resequencing, and further cultured for detailed in vitro and in vivo mechanistic comparison with heterogeneous bulk populations. (B) Clonogenicity. Percentage of single cells which formed colonies under 2D laminin and 3D neurosphere stem cell conditions are given for six pGBM and DIPG primary patient-derived cell cultures, labelled by anatomical location and histone H3 mutation subgroup. (C) 3D neurosphere culture from single cell-derived colonies from SU-DIPG-VI assessed by Celigo S imaging cytometer. (D) Growth of single cell-derived colonies over time, assessed as diammeter of neurosphere, labelled and colour-coded. (E) Targeted sequencing contingency plot of somatic mutations identified as common to all subclones (blue), shared amongst certain subclones (yellow) and private to individuals (red). (F) 2D laminin culture from single cell-derived colonies from SU-DIPG-VI assessed by Celigo S imaging cytometer. (G) Growth of single cell-derived colonies over time, assessed as diammeter of neurosphere, with subclones taken for later analysis highlighted: A-D10 (fast, purple), A-B8 (intermediate, pink) and A-E6 (slow, violet). (H) Targeted sequencing contingency plot of somatic mutations identified as common to all subclones (blue), shared amongst certain subclones (yellow) and private to individuals (red). Gene names are coloured to highlight private mutations in selected subclones, or common to A-D10 and A-B8 (brown). (I) Growth. Time-course for growth of selected subclones re-plated and grown over 160 hours, highlighting statistically significant differences among subclones and heterogeneous bulk cell populations of SU-DIPG-VI (blue). Representative images at 72 hours are provided from the Celigo S cytometer, with tumour cells marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. (J) Invasion. Time-course of invasion of cells into a matrigel matrix over 72 hours, either as percentage of the total area in the field of view covered by invading cells, or as a percentage of time zero. Representative images given at 72 hours, with extent of tumour cell invasion marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. (K) Migration. Time-course of tumour cell migration onto matrigel over 72 hours, either as percentage of the total area of the well covered by migrating cells, or as a percentage of time zero. Representative images given at 72 hours, with extent of tumour cell migration marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. * p

    Journal: Nature medicine

    Article Title: Functional diversity and co-operativity between subclonal populations of paediatric glioblastoma and diffuse intrinsic pontine glioma cells

    doi: 10.1038/s41591-018-0086-7

    Figure Lengend Snippet: Isolation of genotypically and phenotypically diverse single stem-like cell-derived subclones of paediatric GBM and DIPG. (A) Isolation of subclonal populations. Experimental schema for disaggregation of heterogeneous mixtures of patient-derived tumour cells, flow sorting into single cells in 96-well plates, and allowing colonies to form as either 2D cultures adherent on laminin, or 3D neurospheres, all under stem cell conditions. Individual subclonal colonies are subjected to high-throughput phenotypic analysis and targeted resequencing, and further cultured for detailed in vitro and in vivo mechanistic comparison with heterogeneous bulk populations. (B) Clonogenicity. Percentage of single cells which formed colonies under 2D laminin and 3D neurosphere stem cell conditions are given for six pGBM and DIPG primary patient-derived cell cultures, labelled by anatomical location and histone H3 mutation subgroup. (C) 3D neurosphere culture from single cell-derived colonies from SU-DIPG-VI assessed by Celigo S imaging cytometer. (D) Growth of single cell-derived colonies over time, assessed as diammeter of neurosphere, labelled and colour-coded. (E) Targeted sequencing contingency plot of somatic mutations identified as common to all subclones (blue), shared amongst certain subclones (yellow) and private to individuals (red). (F) 2D laminin culture from single cell-derived colonies from SU-DIPG-VI assessed by Celigo S imaging cytometer. (G) Growth of single cell-derived colonies over time, assessed as diammeter of neurosphere, with subclones taken for later analysis highlighted: A-D10 (fast, purple), A-B8 (intermediate, pink) and A-E6 (slow, violet). (H) Targeted sequencing contingency plot of somatic mutations identified as common to all subclones (blue), shared amongst certain subclones (yellow) and private to individuals (red). Gene names are coloured to highlight private mutations in selected subclones, or common to A-D10 and A-B8 (brown). (I) Growth. Time-course for growth of selected subclones re-plated and grown over 160 hours, highlighting statistically significant differences among subclones and heterogeneous bulk cell populations of SU-DIPG-VI (blue). Representative images at 72 hours are provided from the Celigo S cytometer, with tumour cells marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. (J) Invasion. Time-course of invasion of cells into a matrigel matrix over 72 hours, either as percentage of the total area in the field of view covered by invading cells, or as a percentage of time zero. Representative images given at 72 hours, with extent of tumour cell invasion marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. (K) Migration. Time-course of tumour cell migration onto matrigel over 72 hours, either as percentage of the total area of the well covered by migrating cells, or as a percentage of time zero. Representative images given at 72 hours, with extent of tumour cell migration marked in green. Data derived and representative images taken from n=3 independent experiments. Scale bar = 500μm. * p

    Article Snippet: Two 96-well flat bottom plates (Greiner Bio-one) were collected for 2D adherent culture and one 96 well round bottom ultra low attachment plate (Corning) was collected for 3D neurosphere culture.

    Techniques: Isolation, Derivative Assay, Flow Cytometry, High Throughput Screening Assay, Cell Culture, In Vitro, In Vivo, Mutagenesis, Imaging, Cytometry, Sequencing, Migration

    Spreading and migration of VSOP/Hv1 −/− neutrophils . Bone marrow neutrophils were seeded on BSA-coated Greiner 96-well plates and fMIVIL was added to promote chemokinesis. (A) Representative phase-contrast images of neutrophils before (top) and after (bottom) stimulation with fMIVIL+PMA. The combination of fMIVIL and PMA induced 100% of the cells to spread. (B) Percentage of spread cells among naive WT and VSOP/Hv1 −/− neutrophils. Data are means ± SD of 107 WT and 94 VSOP/Hv1 −/− neutrophils from three independent experiments. **, P

    Journal: The Journal of Experimental Medicine

    Article Title: VSOP/Hv1 proton channels sustain calcium entry, neutrophil migration, and superoxide production by limiting cell depolarization and acidification

    doi: 10.1084/jem.20091837

    Figure Lengend Snippet: Spreading and migration of VSOP/Hv1 −/− neutrophils . Bone marrow neutrophils were seeded on BSA-coated Greiner 96-well plates and fMIVIL was added to promote chemokinesis. (A) Representative phase-contrast images of neutrophils before (top) and after (bottom) stimulation with fMIVIL+PMA. The combination of fMIVIL and PMA induced 100% of the cells to spread. (B) Percentage of spread cells among naive WT and VSOP/Hv1 −/− neutrophils. Data are means ± SD of 107 WT and 94 VSOP/Hv1 −/− neutrophils from three independent experiments. **, P

    Article Snippet: Bone marrow neutrophils were seeded in a Greiner 96-well flat-bottom plate at 500,000 cells/well in HBSS medium supplemented with 0.5% BSA, 1% glucose, and 20 mM Hepes.

    Techniques: Migration

    In vitro characterization of the inhibitory activity of the leading compound 9029936 on C. albicans biofilm formation. (A) Dose-dependent inhibitory effects of compound 9029936 on C. albicans biofilm formation. The compound was tested in serial 2-fold dilutions (concentrations ranging from 40 to 0.078 µM), with appropriate positive and negative controls. Results shown are the mean percent biofilm inhibition relative to control biofilms (grown in the absence of compound 9029936), determined in XTT colorimetric assays for multiple technical replicates from several independent experiments. Error bars indicate standard deviations. (B) The leading compound inhibited the proliferation and maturation phases of C. albicans biofilm development. A biofilm kinetic assay was performed in order to examine the inhibitory effects of compound 9029936 at different stages of biofilm development, using the same 96-well microtiter plate model of C. albicans biofilm formation. The extent of inhibition was determined at multiple times (2, 6, 8, 12, 24, and 48 h) after seeding the wells with C. albicans cells in the presence or absence of compound 9029936 (5 μM concentration). Results are expressed as means of multiple technical replicates from a single experiment, with error bars representing standard deviations. (C) SEM images of biofilms of C. albicans strain SC5314 formed in the absence or presence of compound 9029936 at a concentration of 5 μM. Bars, 20 μm.

    Journal: mBio

    Article Title: Development of Anti-Virulence Approaches for Candidiasis via a Novel Series of Small-Molecule Inhibitors of Candida albicans Filamentation

    doi: 10.1128/mBio.01991-17

    Figure Lengend Snippet: In vitro characterization of the inhibitory activity of the leading compound 9029936 on C. albicans biofilm formation. (A) Dose-dependent inhibitory effects of compound 9029936 on C. albicans biofilm formation. The compound was tested in serial 2-fold dilutions (concentrations ranging from 40 to 0.078 µM), with appropriate positive and negative controls. Results shown are the mean percent biofilm inhibition relative to control biofilms (grown in the absence of compound 9029936), determined in XTT colorimetric assays for multiple technical replicates from several independent experiments. Error bars indicate standard deviations. (B) The leading compound inhibited the proliferation and maturation phases of C. albicans biofilm development. A biofilm kinetic assay was performed in order to examine the inhibitory effects of compound 9029936 at different stages of biofilm development, using the same 96-well microtiter plate model of C. albicans biofilm formation. The extent of inhibition was determined at multiple times (2, 6, 8, 12, 24, and 48 h) after seeding the wells with C. albicans cells in the presence or absence of compound 9029936 (5 μM concentration). Results are expressed as means of multiple technical replicates from a single experiment, with error bars representing standard deviations. (C) SEM images of biofilms of C. albicans strain SC5314 formed in the absence or presence of compound 9029936 at a concentration of 5 μM. Bars, 20 μm.

    Article Snippet: For inhibition of biofilm formation, serial 2-fold dilutions (ranging from 40 to 0.078 µM) of compound 9029936 or 7977044 were added to the wells of 96-well flat-bottom microtiter plates before seeding with aliquots of a C. albicans SC5314 culture in RPMI 1640 medium supplemented with l -glutamine (Corning) and buffered with 165 mM morpholinepropanesulfonic acid (MOPS; Sigma; 100 µl/well of a 1 × 106 cells/ml solution).

    Techniques: In Vitro, Activity Assay, Inhibition, Kinetic Assay, Concentration Assay

    Primary screen to identify inhibitors of C. albicans filamentation. (A) Schematic diagram of the phenotypic assay used for large-scale phenotypic screening of 30,000 small-molecule compounds in the DIVERSet chemical library, which we used in our search for inhibitors of C. albicans filamentation. The screening uses 96-well round-bottom microtiter plates and takes advantage of tight control via doxycycline of morphogenetic conversions in the C. albicans tet-NRG1 strain. Individual wells of the microtiter plates are seeded with fungal cells in the presence of 5 µM each individual compound, with appropriate positive and negative controls. The plates are incubated at 37°C and visually inspected at 2 h, 4 h, and 24 h. Under the conditions used, wells containing cells that grow the filamentous form (uninhibited by the presence of the compound) appear cloudy, whereas cells that grow in the yeast form (due to inhibition of filamentation in the presence of a hit compound) fall to the bottom of the wells and form rings, which are easily discernible macroscopically. Microscopy is then used for confirmation of the inhibitory effect on filamentation. (B) Chemical structures of compounds 9029936 and 7977044, two of the major initial hits identified in the primary screen. The two compounds share a common biaryl amide motif (highlighted in red). (C) Identity, physicochemical properties, including the clogP (partition coefficient and a measure of lipophilicity), tPSA (molecular polar surface area), and logSw (solubility of the drug in water), as well as IC 50 (potency) and CC 50 (toxicity) values, for these two small-molecule compounds.

    Journal: mBio

    Article Title: Development of Anti-Virulence Approaches for Candidiasis via a Novel Series of Small-Molecule Inhibitors of Candida albicans Filamentation

    doi: 10.1128/mBio.01991-17

    Figure Lengend Snippet: Primary screen to identify inhibitors of C. albicans filamentation. (A) Schematic diagram of the phenotypic assay used for large-scale phenotypic screening of 30,000 small-molecule compounds in the DIVERSet chemical library, which we used in our search for inhibitors of C. albicans filamentation. The screening uses 96-well round-bottom microtiter plates and takes advantage of tight control via doxycycline of morphogenetic conversions in the C. albicans tet-NRG1 strain. Individual wells of the microtiter plates are seeded with fungal cells in the presence of 5 µM each individual compound, with appropriate positive and negative controls. The plates are incubated at 37°C and visually inspected at 2 h, 4 h, and 24 h. Under the conditions used, wells containing cells that grow the filamentous form (uninhibited by the presence of the compound) appear cloudy, whereas cells that grow in the yeast form (due to inhibition of filamentation in the presence of a hit compound) fall to the bottom of the wells and form rings, which are easily discernible macroscopically. Microscopy is then used for confirmation of the inhibitory effect on filamentation. (B) Chemical structures of compounds 9029936 and 7977044, two of the major initial hits identified in the primary screen. The two compounds share a common biaryl amide motif (highlighted in red). (C) Identity, physicochemical properties, including the clogP (partition coefficient and a measure of lipophilicity), tPSA (molecular polar surface area), and logSw (solubility of the drug in water), as well as IC 50 (potency) and CC 50 (toxicity) values, for these two small-molecule compounds.

    Article Snippet: For inhibition of biofilm formation, serial 2-fold dilutions (ranging from 40 to 0.078 µM) of compound 9029936 or 7977044 were added to the wells of 96-well flat-bottom microtiter plates before seeding with aliquots of a C. albicans SC5314 culture in RPMI 1640 medium supplemented with l -glutamine (Corning) and buffered with 165 mM morpholinepropanesulfonic acid (MOPS; Sigma; 100 µl/well of a 1 × 106 cells/ml solution).

    Techniques: Phenotypic Assay, Incubation, Inhibition, Microscopy, Solubility

    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