nunc 96 well plate  (Thermo Fisher)


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    Structured Review

    Thermo Fisher nunc 96 well plate
    Nunc 96 Well Plate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nunc 96 well plate/product/Thermo Fisher
    Average 99 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    nunc 96 well plate - by Bioz Stars, 2020-03
    99/100 stars

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    Related Articles

    Diagnostic Assay:

    Article Title: The Immunomodulatory Drug Glatiramer Acetate is Also an Effective Antimicrobial Agent that Kills Gram-negative Bacteria
    Article Snippet: Susceptibility to GA antimicrobial effects tested by OASTS E . coli and S . aureus laboratory strains were grown overnight in Sensititre® cation-adjusted Mueller-Hinton broth with N -tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid buffer (CAMHBT, TREK Diagnostic System, Thermo Fisher Scientific, Oakwood village, OH) at 37 °C. .. Growth was analysed in Nunc Edge™ 96-well plates (Thermo Fisher) by OASTS using the oCelloScope™ instrument (Philips BioCell A/S, Allerød, Denmark).

    Centrifugation:

    Article Title: Therapeutic surfactant-stripped frozen micelles
    Article Snippet: The supernatants were transferred to 200 μl DCM, shaken for 5 min for extraction to remove absorbance interference by VK1, followed by subsequent centrifugation at 2,000g for 5 min. Haemolysis percentages were determined by absorbance measurements of the water phase at 540 nm. .. SRBCs (Innovative Research, Inc. # IR1-020ND) containing 5 × 106 cells in 0.1 ml PBS were added to each well of a flat bottom 96-well ELISA microtiter plates (Thermo Scientific # 267312) and the plates were incubated overnight at 4 °C.

    Luciferase:

    Article Title: Pre-exposure Prophylaxis With OspA-Specific Human Monoclonal Antibodies Protects Mice Against Tick Transmission of Lyme Disease Spirochetes
    Article Snippet: Serial dilutions of antibodies were made in 100 µL of BSK-H medium containing 10% of guinea pig complement (Sigma) in a Nunc Edge 96-well plate (Thermo Scientific). .. The spirochete viability was quantified by luciferase detection with Bac-Titer Glo reagent (Promega) and read in a Victor3 multilabel counter (Applied Biosystems).

    Filtration:

    Article Title: Large-Scale Recombinant Expression and Purification of Human Tyrosinase Suitable for Structural Studies
    Article Snippet: Paragraph title: Analytical gel filtration and activity assay ... TYR enzyme activity assays were performed at 298 K using 3,4-dihydroxyphenylalanine (L-DOPA) as substrate in a Nunc™ Edge 96-Well plate (Thermo Fisher Scientific) [ , ].

    Neutralization:

    Article Title: Persistence of the protective immunity and kinetics of the isotype specific antibody response against the viral nucleocapsid protein after experimental Schmallenberg virus infection of sheep
    Article Snippet: Serology Virus neutralization test was carried out as previously described [ ] with the exception that all sera were analysed both without (VNTw/d) and with (VNTd) heat-inactivation of the complement system (30 min at 56 °C) before testing. .. Briefly, successive two fold dilutions of 50 µL of sera, from 1/2 to 1/256 onwards, were made in 50 µL of Dulbecco’s Modified Eagles Medium (Gibco, Life Technologies, Ghent, Belgium) supplemented by 1000 IU penicillin/mL, 50 µg/mL gentamicin (Gibco, Life Technologies, Ghent, Belgium) and 250 µg/mL amphotericin B (Gibco, Life Technologies, Ghent, Belgium) (DMEM) in Nunc Edge 96-well plates (Thermo scientific, Waltham, MA, USA).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Therapeutic surfactant-stripped frozen micelles
    Article Snippet: .. SRBCs (Innovative Research, Inc. # IR1-020ND) containing 5 × 106 cells in 0.1 ml PBS were added to each well of a flat bottom 96-well ELISA microtiter plates (Thermo Scientific # 267312) and the plates were incubated overnight at 4 °C. .. 20 μl of 1.8% glutaraldehyde (AMRESCO, # 0904C089) was gently added to wells containing SRBCs and the plate was incubated for 3 min at 25 °C.

    Article Title: Autocrine Sonic Hedgehog Attenuates Inflammation in Cerulein-Induced Acute Pancreatitis in Mice via Upregulation of IL-10
    Article Snippet: Paragraph title: Enzyme-linked Immunosorbent Assay (ELISA) ... Briefly, a NUNC 96-well plate (Thermo Scientific, MA) was coated with 5 µg/ml of anti-Shh (Sigma Aldrich, MO) or anti-IL-10 (R & D, MN) capture antibody at 4°C overnight, washed with 0.05% PBST and blocked with 5% bovine serum albumin at room temperature for 1 hours, and then incubated with culture supernatants at room temperature for 2 hours.

    Incubation:

    Article Title: Therapeutic surfactant-stripped frozen micelles
    Article Snippet: .. SRBCs (Innovative Research, Inc. # IR1-020ND) containing 5 × 106 cells in 0.1 ml PBS were added to each well of a flat bottom 96-well ELISA microtiter plates (Thermo Scientific # 267312) and the plates were incubated overnight at 4 °C. .. 20 μl of 1.8% glutaraldehyde (AMRESCO, # 0904C089) was gently added to wells containing SRBCs and the plate was incubated for 3 min at 25 °C.

    Article Title: Autocrine Sonic Hedgehog Attenuates Inflammation in Cerulein-Induced Acute Pancreatitis in Mice via Upregulation of IL-10
    Article Snippet: .. Briefly, a NUNC 96-well plate (Thermo Scientific, MA) was coated with 5 µg/ml of anti-Shh (Sigma Aldrich, MO) or anti-IL-10 (R & D, MN) capture antibody at 4°C overnight, washed with 0.05% PBST and blocked with 5% bovine serum albumin at room temperature for 1 hours, and then incubated with culture supernatants at room temperature for 2 hours. .. After 5 times rinsing with 0.05% PBST, 2 µg/ml of detecting antibody was added and incubated at room temperature for 2 hours.

    Article Title: Geometric constraints of endothelial cell migration on electrospun fibres
    Article Snippet: Briefly, 10% (V/V) of PrestoBlue® reagent was added in each well (n = 3) and incubated at 37 °C for 2 hours. .. Three 100 µL media samples were transferred from each well into a Nunc™ 96-well plate (Thermo Scientific, cat.

    Article Title: Persistence of the protective immunity and kinetics of the isotype specific antibody response against the viral nucleocapsid protein after experimental Schmallenberg virus infection of sheep
    Article Snippet: Briefly, successive two fold dilutions of 50 µL of sera, from 1/2 to 1/256 onwards, were made in 50 µL of Dulbecco’s Modified Eagles Medium (Gibco, Life Technologies, Ghent, Belgium) supplemented by 1000 IU penicillin/mL, 50 µg/mL gentamicin (Gibco, Life Technologies, Ghent, Belgium) and 250 µg/mL amphotericin B (Gibco, Life Technologies, Ghent, Belgium) (DMEM) in Nunc Edge 96-well plates (Thermo scientific, Waltham, MA, USA). .. After four days of incubation, the titre was determined as the reciprocal of the highest serum dilution in which no lysis plaques could be identified in the cell monolayer under the light microscope.

    Article Title: Establishing quantitative real-time quaking-induced conversion (qRT-QuIC) for highly sensitive detection and quantification of PrPSc in prion-infected tissues
    Article Snippet: Quantitative RT-QuIC Preparations for quantitative RT-QuIC were loaded into a Nunc 96-well plate (Thermo Scientific, USA) in a bio-safety cabinet. .. Reactions were performed on a FLUOstar Optima (BMG Labtech, Germany) at 37°C for 90 hours with 1 min shaking at 600 rpm followed by 1 min stationary incubation.

    Article Title: Pre-exposure Prophylaxis With OspA-Specific Human Monoclonal Antibodies Protects Mice Against Tick Transmission of Lyme Disease Spirochetes
    Article Snippet: Serial dilutions of antibodies were made in 100 µL of BSK-H medium containing 10% of guinea pig complement (Sigma) in a Nunc Edge 96-well plate (Thermo Scientific). .. Serial dilutions of antibodies were made in 100 µL of BSK-H medium containing 10% of guinea pig complement (Sigma) in a Nunc Edge 96-well plate (Thermo Scientific).

    Article Title: Schwann cells promote endothelial cell migration
    Article Snippet: Briefly, 10% (v/v) of PrestoBlue® reagent was added in each well and the samples (n = 3) incubated at 37 ˚C for 2 hours. .. Three 100 µL media samples were taken from each well into a Nunc™ 96-well plate (Thermo Scientific, cat.

    Article Title: Tuwongella immobilis gen. nov., sp. nov., a novel non-motile bacterium within the phylum Planctomycetes
    Article Snippet: MBLW1T was grown for 48 h at 32 °C in Nunc Edge 96-well microplates (Thermo Scientific) and the optical density (λ=600 nm) was measured using a Tecan Infinite M200 microplate reader (Tecan). .. Salt tolerance of MBLW1T and G. obscuriglobus was compared in 24-well M1 agar plates containing increasing NaCl-concentrations [0.0 %–2.0 % (w/v)] after incubation for 5 days at 32 °C.

    Article Title: Assessment of Cr(VI)-Induced Cytotoxicity and Genotoxicity Using High Content Analysis
    Article Snippet: .. For assays in undifferentiated/proliferating Caco-2 cells, cells were seeded at a density of 1×104 cells/100 µl/well in Nunc Edge 96-well microplates (Thermo Fisher Scientific) 24 hr prior to incubation with compounds. .. For assays in differentiated Caco-2 monolayers, cells were seeded at a density of 2×104 cells/100 µl/well in Collagen I (BD Biosciences) coated Nunc Edge 96-well microplates (Thermo Fisher Scientific), and cultured for 21 days prior to incubation with compounds, with a media change every other day.

    Activity Assay:

    Article Title: Large-Scale Recombinant Expression and Purification of Human Tyrosinase Suitable for Structural Studies
    Article Snippet: .. TYR enzyme activity assays were performed at 298 K using 3,4-dihydroxyphenylalanine (L-DOPA) as substrate in a Nunc™ Edge 96-Well plate (Thermo Fisher Scientific) [ , ]. ..

    Modification:

    Article Title: Persistence of the protective immunity and kinetics of the isotype specific antibody response against the viral nucleocapsid protein after experimental Schmallenberg virus infection of sheep
    Article Snippet: .. Briefly, successive two fold dilutions of 50 µL of sera, from 1/2 to 1/256 onwards, were made in 50 µL of Dulbecco’s Modified Eagles Medium (Gibco, Life Technologies, Ghent, Belgium) supplemented by 1000 IU penicillin/mL, 50 µg/mL gentamicin (Gibco, Life Technologies, Ghent, Belgium) and 250 µg/mL amphotericin B (Gibco, Life Technologies, Ghent, Belgium) (DMEM) in Nunc Edge 96-well plates (Thermo scientific, Waltham, MA, USA). ..

    Flow Cytometry:

    Article Title: Large-Scale Recombinant Expression and Purification of Human Tyrosinase Suitable for Structural Studies
    Article Snippet: Elution from the gel filtration column was carried out at a flow rate of 0.5 mL/min, fractionation volume of 0.3 mL, and total running time of 40 min. .. TYR enzyme activity assays were performed at 298 K using 3,4-dihydroxyphenylalanine (L-DOPA) as substrate in a Nunc™ Edge 96-Well plate (Thermo Fisher Scientific) [ , ].

    Cell Culture:

    Article Title: Assessment of Cr(VI)-Induced Cytotoxicity and Genotoxicity Using High Content Analysis
    Article Snippet: Paragraph title: Cell Culture ... For assays in undifferentiated/proliferating Caco-2 cells, cells were seeded at a density of 1×104 cells/100 µl/well in Nunc Edge 96-well microplates (Thermo Fisher Scientific) 24 hr prior to incubation with compounds.

    Article Title: Quantitative Analysis of Mitochondrial Morphology and Membrane Potential in Living Cells Using High-Content Imaging, Machine Learning, and Morphological Binning
    Article Snippet: .. Cells of passages 15-25 were cultured to 80% confluency before trypsinization and seeded at a cell density of 8,000 cells per well in 96 well plates (Nunc Edge Plate, Thermo Scientific #167314) supplemented with 5% FCS (fetal calf serum) and edge reservoirs filled with phosphate-buffered saline (to prevent hydration-dependent microplate edge effects) (PBS; Life Technologies #14080-055). .. After cells reached 80% confluency (24 h), cells were washed in PBS, and media was changed to DMEM with 5.5 mM glucose without phenol red (Sigma-Aldrich #D-5030) and supplemented with 1% FCS to induce cell cycle arrest.

    Light Microscopy:

    Article Title: Persistence of the protective immunity and kinetics of the isotype specific antibody response against the viral nucleocapsid protein after experimental Schmallenberg virus infection of sheep
    Article Snippet: Briefly, successive two fold dilutions of 50 µL of sera, from 1/2 to 1/256 onwards, were made in 50 µL of Dulbecco’s Modified Eagles Medium (Gibco, Life Technologies, Ghent, Belgium) supplemented by 1000 IU penicillin/mL, 50 µg/mL gentamicin (Gibco, Life Technologies, Ghent, Belgium) and 250 µg/mL amphotericin B (Gibco, Life Technologies, Ghent, Belgium) (DMEM) in Nunc Edge 96-well plates (Thermo scientific, Waltham, MA, USA). .. After four days of incubation, the titre was determined as the reciprocal of the highest serum dilution in which no lysis plaques could be identified in the cell monolayer under the light microscope.

    Injection:

    Article Title: Therapeutic surfactant-stripped frozen micelles
    Article Snippet: SRBCs (Innovative Research, Inc. # IR1-020ND) containing 5 × 106 cells in 0.1 ml PBS were added to each well of a flat bottom 96-well ELISA microtiter plates (Thermo Scientific # 267312) and the plates were incubated overnight at 4 °C. .. Mice were injected intraperitoneally with 5 × 108 SRBCs in 0.5 ml saline.

    Fluorescence:

    Article Title: Geometric constraints of endothelial cell migration on electrospun fibres
    Article Snippet: Three 100 µL media samples were transferred from each well into a Nunc™ 96-well plate (Thermo Scientific, cat. .. Fluorescence was measured at 540–570 nm excitation 580–610 nm emission in VICTOR3 ™ 1420 Multilabel Counter (PerkinElmer).

    Article Title: Schwann cells promote endothelial cell migration
    Article Snippet: Three 100 µL media samples were taken from each well into a Nunc™ 96-well plate (Thermo Scientific, cat. .. Fluorescence was measured at 540–570 nm excitation 580–610 nm emission in VICTOR3 ™ 1420 Multilabel Counter (PerkinElmer).

    Article Title: The combined use of paclitaxel-loaded nanoparticles with a low-molecular-weight copolymer inhibitor of P-glycoprotein to overcome drug resistance
    Article Snippet: .. A small amount of the labeled copolymers was dissolved in DMF, and the TMRCA labeling efficiency of the diblock copolymers was determined by spectrofluorometry against a calibration curve of free TMRCA in DMF using a Synergy MX (Biotek, Winooski, VT, USA) fluorescence plate reader and a Nunc 96-well plate (Thermo Fisher Scientific) using an excitation wavelength of 548 nm and an emission wavelength of 572 nm. .. The fluorescent labeling efficiency for PCL19 and PCL104 was approximately 50.3% and 63.3%, respectively.

    Temperature Regulation:

    Article Title: The Immunomodulatory Drug Glatiramer Acetate is Also an Effective Antimicrobial Agent that Kills Gram-negative Bacteria
    Article Snippet: Growth was analysed in Nunc Edge™ 96-well plates (Thermo Fisher) by OASTS using the oCelloScope™ instrument (Philips BioCell A/S, Allerød, Denmark). .. Each well was scanned repeatedly every 15 min for the first 2 h and every 10 min for the following 5 h. The OASTS instrument was placed inside an Innova 44 incubator (Eppendorf, Hamburg, Germany) for precise temperature regulation.

    Microscopy:

    Article Title: Growth and single cell kinetics of the loricate choanoflagellate Diaphanoeca grandis
    Article Snippet: Single cell growth kinetics Growth of Diaphanoeca grandis was also monitored at the single-cell level in the oCelloScope optical detection system (Biosense Solutions Aps, Denmark), which is an automatic digital microscope equipped with a 4x optical magnification (200x effective magnification) , . .. In the oCelloScope, D. grandis was grown in NUNCTM Edge 96-well plates (Thermo Scientific, USA) containing 200 μL of sterilised seawater (salinity = 10‰, temp.

    Mouse Assay:

    Article Title: Therapeutic surfactant-stripped frozen micelles
    Article Snippet: SRBCs (Innovative Research, Inc. # IR1-020ND) containing 5 × 106 cells in 0.1 ml PBS were added to each well of a flat bottom 96-well ELISA microtiter plates (Thermo Scientific # 267312) and the plates were incubated overnight at 4 °C. .. Mice were injected intraperitoneally with 5 × 108 SRBCs in 0.5 ml saline.

    Labeling:

    Article Title: The combined use of paclitaxel-loaded nanoparticles with a low-molecular-weight copolymer inhibitor of P-glycoprotein to overcome drug resistance
    Article Snippet: .. A small amount of the labeled copolymers was dissolved in DMF, and the TMRCA labeling efficiency of the diblock copolymers was determined by spectrofluorometry against a calibration curve of free TMRCA in DMF using a Synergy MX (Biotek, Winooski, VT, USA) fluorescence plate reader and a Nunc 96-well plate (Thermo Fisher Scientific) using an excitation wavelength of 548 nm and an emission wavelength of 572 nm. .. The fluorescent labeling efficiency for PCL19 and PCL104 was approximately 50.3% and 63.3%, respectively.

    Viability Assay:

    Article Title: Geometric constraints of endothelial cell migration on electrospun fibres
    Article Snippet: Paragraph title: Viability assay ... Three 100 µL media samples were transferred from each well into a Nunc™ 96-well plate (Thermo Scientific, cat.

    Article Title: Schwann cells promote endothelial cell migration
    Article Snippet: Paragraph title: Viability assay ... Three 100 µL media samples were taken from each well into a Nunc™ 96-well plate (Thermo Scientific, cat.

    Time-lapse Microscopy:

    Article Title: The Immunomodulatory Drug Glatiramer Acetate is Also an Effective Antimicrobial Agent that Kills Gram-negative Bacteria
    Article Snippet: Growth was analysed in Nunc Edge™ 96-well plates (Thermo Fisher) by OASTS using the oCelloScope™ instrument (Philips BioCell A/S, Allerød, Denmark). .. OASTS is a digital time-lapse microscopy technology that scans through a fluid sample, generating time-resolved series of images .

    Evaporation:

    Article Title: Growth and single cell kinetics of the loricate choanoflagellate Diaphanoeca grandis
    Article Snippet: In the oCelloScope, D. grandis was grown in NUNCTM Edge 96-well plates (Thermo Scientific, USA) containing 200 μL of sterilised seawater (salinity = 10‰, temp. .. To minimize evaporation, 8 mL of water was added to the surrounding plate-reservoir.

    Concentration Assay:

    Article Title: Autocrine Sonic Hedgehog Attenuates Inflammation in Cerulein-Induced Acute Pancreatitis in Mice via Upregulation of IL-10
    Article Snippet: Briefly, a NUNC 96-well plate (Thermo Scientific, MA) was coated with 5 µg/ml of anti-Shh (Sigma Aldrich, MO) or anti-IL-10 (R & D, MN) capture antibody at 4°C overnight, washed with 0.05% PBST and blocked with 5% bovine serum albumin at room temperature for 1 hours, and then incubated with culture supernatants at room temperature for 2 hours. .. Following 5 times PBST rinsing, the Biotin-HRP conjugated second antibody was added at a concentration of 1∶5000 at room temperature for 1 hour.

    Article Title: Pre-exposure Prophylaxis With OspA-Specific Human Monoclonal Antibodies Protects Mice Against Tick Transmission of Lyme Disease Spirochetes
    Article Snippet: Serial dilutions of antibodies were made in 100 µL of BSK-H medium containing 10% of guinea pig complement (Sigma) in a Nunc Edge 96-well plate (Thermo Scientific). .. A total of 100 µL of Borrelia culture ( B. burgdorferi B31 [ATCC35210], B. garinii PBi [ATCCBAA-2496], and B. afzelii BO23 [ATCC51992]) at a concentration of 5 × 106 spirochetes/mL was added to each well to mix with antibodies.

    Article Title: Assessment of Cr(VI)-Induced Cytotoxicity and Genotoxicity Using High Content Analysis
    Article Snippet: For assays in undifferentiated/proliferating Caco-2 cells, cells were seeded at a density of 1×104 cells/100 µl/well in Nunc Edge 96-well microplates (Thermo Fisher Scientific) 24 hr prior to incubation with compounds. .. Stock solutions of SDD and hydrogen peroxide were prepared in H2 O. Rotenone was prepared in DMSO at a final concentration of 0.25% DMSO.

    Fractionation:

    Article Title: Large-Scale Recombinant Expression and Purification of Human Tyrosinase Suitable for Structural Studies
    Article Snippet: Elution from the gel filtration column was carried out at a flow rate of 0.5 mL/min, fractionation volume of 0.3 mL, and total running time of 40 min. .. TYR enzyme activity assays were performed at 298 K using 3,4-dihydroxyphenylalanine (L-DOPA) as substrate in a Nunc™ Edge 96-Well plate (Thermo Fisher Scientific) [ , ].

    BAC Assay:

    Article Title: Pre-exposure Prophylaxis With OspA-Specific Human Monoclonal Antibodies Protects Mice Against Tick Transmission of Lyme Disease Spirochetes
    Article Snippet: Paragraph title: Borreliacidal Assay by Bac-Titer Glo Detection ... Serial dilutions of antibodies were made in 100 µL of BSK-H medium containing 10% of guinea pig complement (Sigma) in a Nunc Edge 96-well plate (Thermo Scientific).

    Prestoblue Assay:

    Article Title: Geometric constraints of endothelial cell migration on electrospun fibres
    Article Snippet: Cell viability was assessed using PrestoBlue® assay according to the manufacturer’s protocol (Life Technologies, cat. .. Three 100 µL media samples were transferred from each well into a Nunc™ 96-well plate (Thermo Scientific, cat.

    Article Title: Schwann cells promote endothelial cell migration
    Article Snippet: Cell viability was assessed using PrestoBlue® assay according to the manufacturer's protocol (Life Technologies, cat. .. Three 100 µL media samples were taken from each well into a Nunc™ 96-well plate (Thermo Scientific, cat.

    Lysis:

    Article Title: Persistence of the protective immunity and kinetics of the isotype specific antibody response against the viral nucleocapsid protein after experimental Schmallenberg virus infection of sheep
    Article Snippet: Briefly, successive two fold dilutions of 50 µL of sera, from 1/2 to 1/256 onwards, were made in 50 µL of Dulbecco’s Modified Eagles Medium (Gibco, Life Technologies, Ghent, Belgium) supplemented by 1000 IU penicillin/mL, 50 µg/mL gentamicin (Gibco, Life Technologies, Ghent, Belgium) and 250 µg/mL amphotericin B (Gibco, Life Technologies, Ghent, Belgium) (DMEM) in Nunc Edge 96-well plates (Thermo scientific, Waltham, MA, USA). .. After four days of incubation, the titre was determined as the reciprocal of the highest serum dilution in which no lysis plaques could be identified in the cell monolayer under the light microscope.

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  • 99
    Thermo Fisher glass bottom 96 well plates
    HCV triggers stress granule (SG) formation in infected cells. (A) Persistently HCV-infected (at an MOI of 0.1 for 3 weeks) and uninfected control (Mock) Huh-7 cells were seeded in glass bottom 96-well plates, fixed with 4% PFA, and processed for immunofluorescence
    Glass Bottom 96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glass bottom 96 well plates/product/Thermo Fisher
    Average 99 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    glass bottom 96 well plates - by Bioz Stars, 2020-03
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher elisa plates
    Description and characterization of the chimeric human <t>FasL-derived</t> constructs Panel A: Schematic representation of soluble FasL (sFasL), Flag-tagged sFasL (sfFasL), polymeric Flag-tagged soluble FasL (pfFasL), polymeric TCR γ4 and δ5 Flag-tagged soluble FasL generating the TCR-pfFasL upon cotransfection, and beta2-microglobulin-fused HLA-A*02: 01 Flag-tagged soluble FasL (HLA-pfFasL). The f and p symbols represent the flag epitope and the LIF receptor-derived domain triggering the polymerisation of the FasL oligomers, respectively. Panel B: direct immunoblot of the supernatants from COS cells transfected with the empty vector (control) or the FasL constructs sFasL, sfFasL and pfFasL. Panel C: immunoprecipitation of the TCR-pfFasL chimera from transfected HEK cells, using an irrelevant IgG1 antibody, the anti-Flag (clone M2), the anti-FasL (clone 10F2), the anti-TCRγδ (clone IMU-510) or the anti-TCRδ5 (clone 12C7) antibodies. Panel D: immunoprecipitation of the HLA-pfFasL chimera from the supernatant of COS cells, with anti-Flag, anti-FasL or anti-β2microglobulin antibodies. As controls, the same experiment was performed with irrelevant IgG1 and IgG2 antibodies. Panel E: cytotoxic effect of the FasL chimeras. The indicated chimeras, as supernatants from transfected cells and quantitated using the <t>ELISA</t> for FasL, were incubated at the indicated concentrations with Jurkat cells. After 18 h, the MTT cell viability assay was performed. The anti-Flag M2 antibody at 0.5 µg/ml was added to sfFasL to render it cytotoxic.
    Elisa Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 430 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa plates/product/Thermo Fisher
    Average 99 stars, based on 430 article reviews
    Price from $9.99 to $1999.99
    elisa plates - by Bioz Stars, 2020-03
    99/100 stars
      Buy from Supplier

    78
    Thermo Fisher grp78 elisa microtiter 96 well plates
    Analysis of <t>GRP78–HTJ1</t> and OxPL–GRP78 interactions. (A) GRP78 and HTJ1 expression in HPAEC and human lung microvascular endothelial cells was detected by Western Blot. (B, C) GRP78 interactions were analyzed in coimmunoprecipitation assays using lysates from control or DMPC- (10 μg/ml, 15 min) or OxPAPC-stimulated (10 μg/ml) cells with antibody to GRP78 (B, top), HTJ1 (B, bottom), or EO6 antibody recognizing OxPL (C). (D) Human recombinant GRP78 was incubated with OxPAPC, OxPAPS, or their oxidation-resistant analogues DMPC or DMPS. Left, native gel electrophoresis, followed by Western blot with anti-GRP78 antibody. Shift in electrophoretic mobility of GRP78 incubated with OxPAPS, but not DMPS, indicates formation of GRP78–OxPAPS complex. Right, SDS–PAGE, followed by Western blot with EO6 antibody and reprobing with anti-GRP78 antibody. Positive EO6 immunoreactivity of GRP78 preincubated with OxPAPC indicates formation of GRP78–OxPAPC complex. (E) <t>ELISA</t> plates coated with OxPAPS or DMPS or control uncoated plates incubated with PBS incubated with human recombinant GRP78 (left) or HPAEC lysates (middle and right). The bound GRP78 was detected using anti-GRP78 antibody. * p
    Grp78 Elisa Microtiter 96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/grp78 elisa microtiter 96 well plates/product/Thermo Fisher
    Average 78 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    grp78 elisa microtiter 96 well plates - by Bioz Stars, 2020-03
    78/100 stars
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    HCV triggers stress granule (SG) formation in infected cells. (A) Persistently HCV-infected (at an MOI of 0.1 for 3 weeks) and uninfected control (Mock) Huh-7 cells were seeded in glass bottom 96-well plates, fixed with 4% PFA, and processed for immunofluorescence

    Journal: Journal of Virology

    Article Title: Hepatitis C Virus (HCV) Induces Formation of Stress Granules Whose Proteins Regulate HCV RNA Replication and Virus Assembly and Egress

    doi: 10.1128/JVI.07101-11

    Figure Lengend Snippet: HCV triggers stress granule (SG) formation in infected cells. (A) Persistently HCV-infected (at an MOI of 0.1 for 3 weeks) and uninfected control (Mock) Huh-7 cells were seeded in glass bottom 96-well plates, fixed with 4% PFA, and processed for immunofluorescence

    Article Snippet: For confocal immunofluorescence experiments, Huh-7 cells were grown in glass bottom 96-well plates (Nunc; Thermo Scientific, Rochester, NY) and treated or infected as indicated in the figure legends.

    Techniques: Infection, Immunofluorescence

    Effect of azithromycin up to 5.0 mg/L on the red complex mono- and polymicrobial biofilms in a 96-well plate model. Azithromycin at concentrations 0–100 mg/L was incubated with bacterial cultures for 48 h under anaerobic conditions. Data points represent the mean AU 620 value of a minimum of three biological replicates. Note the categorical scale.

    Journal: Journal of Oral Microbiology

    Article Title: Effect of azithromycin on a red complex polymicrobial biofilm

    doi: 10.1080/20002297.2017.1339579

    Figure Lengend Snippet: Effect of azithromycin up to 5.0 mg/L on the red complex mono- and polymicrobial biofilms in a 96-well plate model. Azithromycin at concentrations 0–100 mg/L was incubated with bacterial cultures for 48 h under anaerobic conditions. Data points represent the mean AU 620 value of a minimum of three biological replicates. Note the categorical scale.

    Article Snippet: Two hundred microliters of P. gingivalis , T. denticola , or T. forsythia as a monospecies inoculum and the combination of each two bacterial species at equal volumes (100 µL each), as well as all three species (67 µL each) as a polymicrobial inoculum, were aliquoted into 96-well flat-bottom plates (Nunc; Thermo Scientific) to provide the same total number of bacterial cells per inoculum.

    Techniques: Incubation

    Formation of mono- and polymicrobial biofilms in a 96-well plate model after 48 h of incubation at 37°C under anaerobic condition. Native bacterial growth with addition of uncultured growth medium and no antibiotic served as controls. Adherent biofilms were stained with 0.1% crystal violet and the optical density at AU 620 was measured. Data represent the mean AU 620 value of a minimum of three biological replicates.

    Journal: Journal of Oral Microbiology

    Article Title: Effect of azithromycin on a red complex polymicrobial biofilm

    doi: 10.1080/20002297.2017.1339579

    Figure Lengend Snippet: Formation of mono- and polymicrobial biofilms in a 96-well plate model after 48 h of incubation at 37°C under anaerobic condition. Native bacterial growth with addition of uncultured growth medium and no antibiotic served as controls. Adherent biofilms were stained with 0.1% crystal violet and the optical density at AU 620 was measured. Data represent the mean AU 620 value of a minimum of three biological replicates.

    Article Snippet: Two hundred microliters of P. gingivalis , T. denticola , or T. forsythia as a monospecies inoculum and the combination of each two bacterial species at equal volumes (100 µL each), as well as all three species (67 µL each) as a polymicrobial inoculum, were aliquoted into 96-well flat-bottom plates (Nunc; Thermo Scientific) to provide the same total number of bacterial cells per inoculum.

    Techniques: Incubation, Staining

    Effects of azithromycin and amoxicillin + metronidazole (1:1 ratio) up to 5.0 mg/L on formation polymicrobial biofilms after 48 h of anaerobic incubation at 37°C in a 96-well plate model. Azithromycin and amoxicillin + metronidazole (1:1 ratio) at concentrations 0–100 mg/L were incubated with bacterial cultures. Data points represent the mean AU 620 value of a minimum of three biological replicates and the standard deviation. * p

    Journal: Journal of Oral Microbiology

    Article Title: Effect of azithromycin on a red complex polymicrobial biofilm

    doi: 10.1080/20002297.2017.1339579

    Figure Lengend Snippet: Effects of azithromycin and amoxicillin + metronidazole (1:1 ratio) up to 5.0 mg/L on formation polymicrobial biofilms after 48 h of anaerobic incubation at 37°C in a 96-well plate model. Azithromycin and amoxicillin + metronidazole (1:1 ratio) at concentrations 0–100 mg/L were incubated with bacterial cultures. Data points represent the mean AU 620 value of a minimum of three biological replicates and the standard deviation. * p

    Article Snippet: Two hundred microliters of P. gingivalis , T. denticola , or T. forsythia as a monospecies inoculum and the combination of each two bacterial species at equal volumes (100 µL each), as well as all three species (67 µL each) as a polymicrobial inoculum, were aliquoted into 96-well flat-bottom plates (Nunc; Thermo Scientific) to provide the same total number of bacterial cells per inoculum.

    Techniques: Incubation, Standard Deviation

    Effect of amoxicillin + metronidazole up to 5.0 mg/L on the red complex mono- and polymicrobial biofilms in a 96-well plate model. Amoxicillin + metronidazole in a 1:1 ratio at concentrations 0–100 mg/L was incubated with bacterial cultures for 48 h at 37°C anaerobically. Data points represent the mean AU 620 value of a minimum of three biological replicates. Note the categorical scale.

    Journal: Journal of Oral Microbiology

    Article Title: Effect of azithromycin on a red complex polymicrobial biofilm

    doi: 10.1080/20002297.2017.1339579

    Figure Lengend Snippet: Effect of amoxicillin + metronidazole up to 5.0 mg/L on the red complex mono- and polymicrobial biofilms in a 96-well plate model. Amoxicillin + metronidazole in a 1:1 ratio at concentrations 0–100 mg/L was incubated with bacterial cultures for 48 h at 37°C anaerobically. Data points represent the mean AU 620 value of a minimum of three biological replicates. Note the categorical scale.

    Article Snippet: Two hundred microliters of P. gingivalis , T. denticola , or T. forsythia as a monospecies inoculum and the combination of each two bacterial species at equal volumes (100 µL each), as well as all three species (67 µL each) as a polymicrobial inoculum, were aliquoted into 96-well flat-bottom plates (Nunc; Thermo Scientific) to provide the same total number of bacterial cells per inoculum.

    Techniques: Incubation

    Description and characterization of the chimeric human FasL-derived constructs Panel A: Schematic representation of soluble FasL (sFasL), Flag-tagged sFasL (sfFasL), polymeric Flag-tagged soluble FasL (pfFasL), polymeric TCR γ4 and δ5 Flag-tagged soluble FasL generating the TCR-pfFasL upon cotransfection, and beta2-microglobulin-fused HLA-A*02: 01 Flag-tagged soluble FasL (HLA-pfFasL). The f and p symbols represent the flag epitope and the LIF receptor-derived domain triggering the polymerisation of the FasL oligomers, respectively. Panel B: direct immunoblot of the supernatants from COS cells transfected with the empty vector (control) or the FasL constructs sFasL, sfFasL and pfFasL. Panel C: immunoprecipitation of the TCR-pfFasL chimera from transfected HEK cells, using an irrelevant IgG1 antibody, the anti-Flag (clone M2), the anti-FasL (clone 10F2), the anti-TCRγδ (clone IMU-510) or the anti-TCRδ5 (clone 12C7) antibodies. Panel D: immunoprecipitation of the HLA-pfFasL chimera from the supernatant of COS cells, with anti-Flag, anti-FasL or anti-β2microglobulin antibodies. As controls, the same experiment was performed with irrelevant IgG1 and IgG2 antibodies. Panel E: cytotoxic effect of the FasL chimeras. The indicated chimeras, as supernatants from transfected cells and quantitated using the ELISA for FasL, were incubated at the indicated concentrations with Jurkat cells. After 18 h, the MTT cell viability assay was performed. The anti-Flag M2 antibody at 0.5 µg/ml was added to sfFasL to render it cytotoxic.

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Description and characterization of the chimeric human FasL-derived constructs Panel A: Schematic representation of soluble FasL (sFasL), Flag-tagged sFasL (sfFasL), polymeric Flag-tagged soluble FasL (pfFasL), polymeric TCR γ4 and δ5 Flag-tagged soluble FasL generating the TCR-pfFasL upon cotransfection, and beta2-microglobulin-fused HLA-A*02: 01 Flag-tagged soluble FasL (HLA-pfFasL). The f and p symbols represent the flag epitope and the LIF receptor-derived domain triggering the polymerisation of the FasL oligomers, respectively. Panel B: direct immunoblot of the supernatants from COS cells transfected with the empty vector (control) or the FasL constructs sFasL, sfFasL and pfFasL. Panel C: immunoprecipitation of the TCR-pfFasL chimera from transfected HEK cells, using an irrelevant IgG1 antibody, the anti-Flag (clone M2), the anti-FasL (clone 10F2), the anti-TCRγδ (clone IMU-510) or the anti-TCRδ5 (clone 12C7) antibodies. Panel D: immunoprecipitation of the HLA-pfFasL chimera from the supernatant of COS cells, with anti-Flag, anti-FasL or anti-β2microglobulin antibodies. As controls, the same experiment was performed with irrelevant IgG1 and IgG2 antibodies. Panel E: cytotoxic effect of the FasL chimeras. The indicated chimeras, as supernatants from transfected cells and quantitated using the ELISA for FasL, were incubated at the indicated concentrations with Jurkat cells. After 18 h, the MTT cell viability assay was performed. The anti-Flag M2 antibody at 0.5 µg/ml was added to sfFasL to render it cytotoxic.

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Derivative Assay, Construct, Cotransfection, FLAG-tag, Transfection, Plasmid Preparation, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Incubation, MTT Assay, Viability Assay

    Direct association of sFasL to the pfFasL-containing chimeric proteins during co-expression. Panel A: Identical amounts of pfFasL (1 µg, according to the Flag ELISA) produced in the presence of the indicated ratios of added sFasL plasmid (left panels) was immunoprecipitated with the anti-FasL (upper panel) or anti-Flag (lower panel) antibodies, followed by a SDS-PAGE under reducing conditions and immunoblotting with an anti-FasL antibody. As a control, the same experiment was performed for the sFasL molecule (3 µg according to the FasL ELISA, right panel). Panel B: Densitometric detection and quantification of the pfFasL (grey bars) and the sFasL (black bars) fractions, following transfection of the pfFasL plasmid in the presence of the indicated proportion of the sFasL plasmid. The measures were normalized to the condition lacking sFasL. Mean+/- sd from three experiments. Panel C: The TCR-pfFasL chimera (2 µg, according to an ELISA specific for the TCR-pFasL molecule using anti-TCRδ5 (clone 12C7) and anti-FasL (clone 10F2) as capture and tracing antibodies, respectively), produced in the absence or the presence of the sFasL plasmid at the indicated ratio, was immunoprecipitated with the anti-TCRδ5 antibody, then separated by 10% SDS-PAGE under reducing conditions and revealed by immunoblotting with the anti-FasL antibody. As a control, the immunoprecipitation experiment was performed with 2 µg of sFasL protein. Panel D: COS supernatants containing pfFasL (4 µg/ml according to the Flag ELISA) produced alone, was mixed with culture medium or sFasL (15 µg/ml) produced separately in a total volume of 1 ml, and incubated for 24 h at 37°C. Then the recombinant proteins were immunoprecipitated (left panels) with the anti-FasL (upper panel) or anti-Flag (lower panel) antibodies, followed by a SDS-PAGE under reducing conditions and immunoblotting with an anti-FasL antibody. As a control, the same experiment was performed for the sFasL molecule (15 µg according to the FasL ELISA, right panel).

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Direct association of sFasL to the pfFasL-containing chimeric proteins during co-expression. Panel A: Identical amounts of pfFasL (1 µg, according to the Flag ELISA) produced in the presence of the indicated ratios of added sFasL plasmid (left panels) was immunoprecipitated with the anti-FasL (upper panel) or anti-Flag (lower panel) antibodies, followed by a SDS-PAGE under reducing conditions and immunoblotting with an anti-FasL antibody. As a control, the same experiment was performed for the sFasL molecule (3 µg according to the FasL ELISA, right panel). Panel B: Densitometric detection and quantification of the pfFasL (grey bars) and the sFasL (black bars) fractions, following transfection of the pfFasL plasmid in the presence of the indicated proportion of the sFasL plasmid. The measures were normalized to the condition lacking sFasL. Mean+/- sd from three experiments. Panel C: The TCR-pfFasL chimera (2 µg, according to an ELISA specific for the TCR-pFasL molecule using anti-TCRδ5 (clone 12C7) and anti-FasL (clone 10F2) as capture and tracing antibodies, respectively), produced in the absence or the presence of the sFasL plasmid at the indicated ratio, was immunoprecipitated with the anti-TCRδ5 antibody, then separated by 10% SDS-PAGE under reducing conditions and revealed by immunoblotting with the anti-FasL antibody. As a control, the immunoprecipitation experiment was performed with 2 µg of sFasL protein. Panel D: COS supernatants containing pfFasL (4 µg/ml according to the Flag ELISA) produced alone, was mixed with culture medium or sFasL (15 µg/ml) produced separately in a total volume of 1 ml, and incubated for 24 h at 37°C. Then the recombinant proteins were immunoprecipitated (left panels) with the anti-FasL (upper panel) or anti-Flag (lower panel) antibodies, followed by a SDS-PAGE under reducing conditions and immunoblotting with an anti-FasL antibody. As a control, the same experiment was performed for the sFasL molecule (15 µg according to the FasL ELISA, right panel).

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Produced, Plasmid Preparation, Immunoprecipitation, SDS Page, Transfection, Incubation, Recombinant

    Effect of sFasL on the supernatant production of the Flag-tagged FasL constructs. Panels A to D : An increasing amount expressed in percentage, of the sFasL encoding plasmid, was co-transfected with a fixed amount of the plasmids encoding sfFasL (Panel A), pfFasL (Panel B), TCR-pfFasL (Panel C) and HLA-pfFasL (Panel D). The secreted proteins were quantified in culture supernatants using an ELISA specific for FasL (shaded histograms, right-hand scale) and for Flag-tagged FasL (curves, left-hand scale). For the Flag ELISA, the measured concentrations were normalized according to the condition lacking sFasL. Are presented the mean +/- sd of four independent transfection experiments. * 0.02≤p≤0.05; ** p≤0.02. Panel E : direct anti-FasL immunoblot analysis of identical volumes of the cell culture supernatant containing pfFasL produced alone and with 50% of the sFasL plasmid, after SDS-PAGE separation under reducing conditions.

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Effect of sFasL on the supernatant production of the Flag-tagged FasL constructs. Panels A to D : An increasing amount expressed in percentage, of the sFasL encoding plasmid, was co-transfected with a fixed amount of the plasmids encoding sfFasL (Panel A), pfFasL (Panel B), TCR-pfFasL (Panel C) and HLA-pfFasL (Panel D). The secreted proteins were quantified in culture supernatants using an ELISA specific for FasL (shaded histograms, right-hand scale) and for Flag-tagged FasL (curves, left-hand scale). For the Flag ELISA, the measured concentrations were normalized according to the condition lacking sFasL. Are presented the mean +/- sd of four independent transfection experiments. * 0.02≤p≤0.05; ** p≤0.02. Panel E : direct anti-FasL immunoblot analysis of identical volumes of the cell culture supernatant containing pfFasL produced alone and with 50% of the sFasL plasmid, after SDS-PAGE separation under reducing conditions.

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Construct, Plasmid Preparation, Transfection, Enzyme-linked Immunosorbent Assay, Cell Culture, Produced, SDS Page

    Effect of sFasL on cell targeting of the FasL-containing chimeras. Panel A : Schematic description of the experimental model used. The chimera is enriched at the surface of the CD32-expressing L-cells via its HLA targeting module and an anti-HLA monoclonal antibody. Panel B: murine Fas (continuous line), human CD32 (dashed line) and IgG1 isotype-matched control (shaded histogram) staining of the CD32+ L-cell transfectant. Living cells were gated on the basis of the morphological parameters. Panel C : Fas sensitivity of the CD32+ L-cell transfectant to the indicated concentrations of the anti-Fas JO-2 antibody (circles), the HLA-pfFasL chimera expressed alone (triangle) or in the presence of 25% of the sFasL plasmid (squares), in the MTT viability assay. Panel D : The CD32+ L-cells were incubated with the HLA-pfFasL chimera produced in the presence (black bars) or in the absence (white bars) of 25% of the sFasL plasmid, together with the indicated irrelevant IgG1 isotype-matched, anti-beta-2 microglobulin or anti-Flag antibodies. The concentrations of the chimera that triggered 15% of cell death and were at 15 and 0.3 ng/ml in the absence and presence of sFasL, as estimated using the ELISA specific for the Flag-tagged FasL. Cytotoxicity was measured with the propidium iodide assay and normalized to the effect of the chimera in the absence of antibody. Are presented the mean +/- sd of three independent experiments. Panel E: reversal in the presence of the blocking anti-FasL and anti-CD32 antibodies, of the cytotoxic effect of the immune complexes between the anti-Flag antibody and HLA-pfFasL co-expressed with sFasL. Are presented the mean +/- sd of three independent experiments. ns : non significant ; ** p≤0.02.

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Effect of sFasL on cell targeting of the FasL-containing chimeras. Panel A : Schematic description of the experimental model used. The chimera is enriched at the surface of the CD32-expressing L-cells via its HLA targeting module and an anti-HLA monoclonal antibody. Panel B: murine Fas (continuous line), human CD32 (dashed line) and IgG1 isotype-matched control (shaded histogram) staining of the CD32+ L-cell transfectant. Living cells were gated on the basis of the morphological parameters. Panel C : Fas sensitivity of the CD32+ L-cell transfectant to the indicated concentrations of the anti-Fas JO-2 antibody (circles), the HLA-pfFasL chimera expressed alone (triangle) or in the presence of 25% of the sFasL plasmid (squares), in the MTT viability assay. Panel D : The CD32+ L-cells were incubated with the HLA-pfFasL chimera produced in the presence (black bars) or in the absence (white bars) of 25% of the sFasL plasmid, together with the indicated irrelevant IgG1 isotype-matched, anti-beta-2 microglobulin or anti-Flag antibodies. The concentrations of the chimera that triggered 15% of cell death and were at 15 and 0.3 ng/ml in the absence and presence of sFasL, as estimated using the ELISA specific for the Flag-tagged FasL. Cytotoxicity was measured with the propidium iodide assay and normalized to the effect of the chimera in the absence of antibody. Are presented the mean +/- sd of three independent experiments. Panel E: reversal in the presence of the blocking anti-FasL and anti-CD32 antibodies, of the cytotoxic effect of the immune complexes between the anti-Flag antibody and HLA-pfFasL co-expressed with sFasL. Are presented the mean +/- sd of three independent experiments. ns : non significant ; ** p≤0.02.

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Expressing, Staining, Transfection, Plasmid Preparation, MTT Assay, Viability Assay, Incubation, Produced, Enzyme-linked Immunosorbent Assay, Blocking Assay

    Effect of sFasL on the cytotoxic activity of the Flag-tagged FasL chimeras. The FasL-derived proteins sfFasL (Panel A), pfFasL (Panel B), TCR-pfFasL (Panel C) and HLA-pfFasL (Panel D) were expressed alone or upon co-transfection with the indicated percentage of the plasmid encoding sFasL. A fixed concentration triggering 25 to 40% of cell death (1.9 ng/ml for sfFasL, 0.6 ng/ml for pfFasL, 0.7 ng/ml for HLA-pfFasL and 2.2 ng/ml for TCR-pfFasL), for the FasL-derived protein quantitated with the ELISA specific for Flag-tagged FasL, was incubated with the Fas-sensitive Jurkat cells. For the sfFasL construct, the filled squares and the empty squares depict the cytotoxicity of sfFasL in the presence and absence of the cross-linking anti-Flag antibody at 0.5 µg/ml), respectively. Cytotoxicity was estimated by a measure of the remaining viable cells using the MTT assay. Are presented the mean +/- sd of four independent transfection experiments. * 0.01≤p≤0.05; ** p≤0.01.

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Effect of sFasL on the cytotoxic activity of the Flag-tagged FasL chimeras. The FasL-derived proteins sfFasL (Panel A), pfFasL (Panel B), TCR-pfFasL (Panel C) and HLA-pfFasL (Panel D) were expressed alone or upon co-transfection with the indicated percentage of the plasmid encoding sFasL. A fixed concentration triggering 25 to 40% of cell death (1.9 ng/ml for sfFasL, 0.6 ng/ml for pfFasL, 0.7 ng/ml for HLA-pfFasL and 2.2 ng/ml for TCR-pfFasL), for the FasL-derived protein quantitated with the ELISA specific for Flag-tagged FasL, was incubated with the Fas-sensitive Jurkat cells. For the sfFasL construct, the filled squares and the empty squares depict the cytotoxicity of sfFasL in the presence and absence of the cross-linking anti-Flag antibody at 0.5 µg/ml), respectively. Cytotoxicity was estimated by a measure of the remaining viable cells using the MTT assay. Are presented the mean +/- sd of four independent transfection experiments. * 0.01≤p≤0.05; ** p≤0.01.

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Activity Assay, Derivative Assay, Cotransfection, Plasmid Preparation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Incubation, Construct, MTT Assay, Transfection

    Analysis of GRP78–HTJ1 and OxPL–GRP78 interactions. (A) GRP78 and HTJ1 expression in HPAEC and human lung microvascular endothelial cells was detected by Western Blot. (B, C) GRP78 interactions were analyzed in coimmunoprecipitation assays using lysates from control or DMPC- (10 μg/ml, 15 min) or OxPAPC-stimulated (10 μg/ml) cells with antibody to GRP78 (B, top), HTJ1 (B, bottom), or EO6 antibody recognizing OxPL (C). (D) Human recombinant GRP78 was incubated with OxPAPC, OxPAPS, or their oxidation-resistant analogues DMPC or DMPS. Left, native gel electrophoresis, followed by Western blot with anti-GRP78 antibody. Shift in electrophoretic mobility of GRP78 incubated with OxPAPS, but not DMPS, indicates formation of GRP78–OxPAPS complex. Right, SDS–PAGE, followed by Western blot with EO6 antibody and reprobing with anti-GRP78 antibody. Positive EO6 immunoreactivity of GRP78 preincubated with OxPAPC indicates formation of GRP78–OxPAPC complex. (E) ELISA plates coated with OxPAPS or DMPS or control uncoated plates incubated with PBS incubated with human recombinant GRP78 (left) or HPAEC lysates (middle and right). The bound GRP78 was detected using anti-GRP78 antibody. * p

    Journal: Molecular Biology of the Cell

    Article Title: GRP78 is a novel receptor initiating a vascular barrier protective response to oxidized phospholipids

    doi: 10.1091/mbc.E13-12-0743

    Figure Lengend Snippet: Analysis of GRP78–HTJ1 and OxPL–GRP78 interactions. (A) GRP78 and HTJ1 expression in HPAEC and human lung microvascular endothelial cells was detected by Western Blot. (B, C) GRP78 interactions were analyzed in coimmunoprecipitation assays using lysates from control or DMPC- (10 μg/ml, 15 min) or OxPAPC-stimulated (10 μg/ml) cells with antibody to GRP78 (B, top), HTJ1 (B, bottom), or EO6 antibody recognizing OxPL (C). (D) Human recombinant GRP78 was incubated with OxPAPC, OxPAPS, or their oxidation-resistant analogues DMPC or DMPS. Left, native gel electrophoresis, followed by Western blot with anti-GRP78 antibody. Shift in electrophoretic mobility of GRP78 incubated with OxPAPS, but not DMPS, indicates formation of GRP78–OxPAPS complex. Right, SDS–PAGE, followed by Western blot with EO6 antibody and reprobing with anti-GRP78 antibody. Positive EO6 immunoreactivity of GRP78 preincubated with OxPAPC indicates formation of GRP78–OxPAPC complex. (E) ELISA plates coated with OxPAPS or DMPS or control uncoated plates incubated with PBS incubated with human recombinant GRP78 (left) or HPAEC lysates (middle and right). The bound GRP78 was detected using anti-GRP78 antibody. * p

    Article Snippet: GRP78 ELISA Microtiter 96-well plates (MaxiSorp; Nunc, Thermo Scientific, Rochester, NY) were coated with OxPAPS or DMPS (each 100 μg/ml in PBS containing 0.01% BHT) at 4°C overnight.

    Techniques: Expressing, Western Blot, Recombinant, Incubation, Nucleic Acid Electrophoresis, SDS Page, Enzyme-linked Immunosorbent Assay