escherichia coli novablue competent cells  (Millipore)


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    Structured Review

    Millipore escherichia coli novablue competent cells
    Escherichia Coli Novablue Competent Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/escherichia coli novablue competent cells/product/Millipore
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    escherichia coli novablue competent cells - by Bioz Stars, 2020-04
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    Related Articles

    Clone Assay:

    Article Title: Characterization of Aphanizomenon ovalisporum amidinotransferase involved in cylindrospermopsin synthesis
    Article Snippet: .. Two Escherichia coli strains were utilized: DH5α, for cloning and sequencing the aoa A gene into pET28b+ vector (Novagen, Madison, WI), and BL21 (DE3) used for protein overexpression. .. Cloning of aoaA and overexpression of AoaA Cyanobacteria DNA extraction and purification were carried out following the method developed by Smoker and Barnum , and modified by Neilan ( ).

    Cell Isolation:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: Paragraph title: Adipose Stem Cell Isolation and Culture ... BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: Paragraph title: Cell Isolation and Culture ... The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1.

    Filtration:

    Article Title: Elevated Carbon Monoxide to Carbon Dioxide Ratio in the Exhaled Breath of Mice Treated With a Single Dose of Lipopolysaccharide
    Article Snippet: .. Reagents Escherichia coli 0111:B4 LPS, which had been phenol-extracted and chromatographically purified by gel filtration and then passed through 0.2-µm pore-sized membrane as a 1.0-mg/mL aqueous solution, was from Sigma-Aldrich (catalog L5293). .. The LPS solution or water was further diluted for use in phosphate-buffered saline (pH 7.4) solution in endotoxin-free Milli-Q (Millipore) water.

    Cytometry:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: After primary cell culture (passages 1–2), the surface marker expression of hASCs was analyzed by flow cytometry (fluorescence-activated cell sorting) (FACSAria; Becton, Dickinson and Company, Erembodegem, Belgium, ) ( ) [ ]. .. BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: The purity of CD14+ monocytes and CD3+ T cell populations was usually higher than 90%, as evaluated by flow cytometry analysis (CyFlow Cube 6, Sysmex Partec GmbH, Görlitz, Germany) (data not shown). .. The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1.

    Blocking Assay:

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1. .. To determine the role of toll-like receptors (TLR)2 and TLR4 on the DCs status induced by ES L1, immature DCs were treated with blocking antibodies against TLR2 and/or TLR4 (10 µg/ml each, both from BioLegend), or isotype control antibody (anti-rat IgG, eBioscience), 1 h before the treatment with ES L1.

    Incubation:

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: The corneal fibroblasts were cultured overnight in 25 cm2 flasks, the medium with non‐attached cells was discarded and the incubation was continued in a new medium which was changed every 3 days, until 90% confluence was achieved. .. The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay).

    Activity Assay:

    Article Title: Targeting proteins for degradation
    Article Snippet: .. Bougdour A, Wickner S, Gottesman S. Modulating RssB activity: IraP, a novel regulator of sigma(S) stability in Escherichia coli. ..

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: For the quantitative alkaline phosphatase (qALP) activity, Alizarin red mineralization analyses, and Oil Red O analyses, 500 cells per well were plated on 24-well plates. .. BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    Solubility:

    Article Title: Oleuropein Decreases Cyclooxygenase-2 and Interleukin-17 Expression and Attenuates Inflammatory Damage in Colonic Samples from Ulcerative Colitis Patients
    Article Snippet: Then, the dishes were placed in an organ culture chamber at 37 °C in 95% O2 , 5% CO2 for 4 and 20 h, and biopsies were stimulated or not (untreated controls) with LPS from Escherichia coli (EC-LPS, Escherichia coli Serotype O127:B8 Lipopolysaccharide, Sigma-Aldrich, Milan, Italy) at a concentration of 1 μg/mL, to induce an inflammatory response, in the absence or presence of OLE at a final concentration of 3 mM. .. Due to its high water solubility, OLE was dissolved in the same medium used for organ culture, to give a stock solution at a concentration of 20 mM.

    Expressing:

    Article Title: Novel fusion proteins for the antigen-specific staining and elimination of B cell receptor-positive cell populations demonstrated by a tetanus toxoid fragment C (TTC) model antigen
    Article Snippet: .. Expression of TTC-ETA’ and TTC in Escherichia coli and protein purification Escherichia coli BL21 (DE3) cells (Novagen, Darmstadt, Germany) were transformed with the TTC and TTC-ETA’ encoding expression vectors and the corresponding proteins were expressed into the periplasm under osmotic stress in the presence of compatible solutes [ ]. .. The protein was purified from the periplasmic fraction by immobilized metal-ion affinity chromatography (IMAC) using a Nickel-Sepharose (Ni-NTA) Superflow Cartridge (Qiagen, Hilden, Germany) on the ÄKTApurifier system (GE Healthcare Life Sciences, Freiburg, Germany) followed by a size-exclusion chromatography using a Superdex 200 (GE Healthcare).

    Article Title: Receptor tyrosine kinase‐like orphan receptor 1, a target of NKX2‐1/TTF‐1 lineage‐survival oncogene, inhibits apoptosis signal‐regulating kinase 1‐mediated pro‐apoptotic signaling in lung adenocarcinoma
    Article Snippet: .. An Escherichia coli ‐expressing protein of MBP‐tagged kinase‐dead MKK6 was purchased from Millipore. .. Western blotting and immunoprecipitation–WB analyses Western blotting and immunoprecipitation (IP)‐WB analyses were carried out according to standard procedures using Immobilon‐P filters (Millipore) and an Amersham ECL Western Blotting Detection Reagent (GE Healthcare, Amersham, UK).

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: After primary cell culture (passages 1–2), the surface marker expression of hASCs was analyzed by flow cytometry (fluorescence-activated cell sorting) (FACSAria; Becton, Dickinson and Company, Erembodegem, Belgium, ) ( ) [ ]. .. BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay). .. The total RNA of treated and non‐treated cells was obtained with Trizol reagent (Invitrogen) for analysis, through RT‐PCR, mRNA expression of: α‐defensin‐3 (AD‐3), α‐defensin‐4 (AD‐4), α‐defensin‐5 (AD‐5), β‐defensin‐1 (BD‐1), β‐defensin‐2 (BD‐2), β‐defensin‐3 (BD‐3), cathelin related antimicrobial peptide (CRAMP), TLR‐1, TLR‐2, TLR‐4, TLR‐6, and TLR‐9.

    Article Title: Screen of Non-annotated Small Secreted Proteins of Pseudomonas syringae Reveals a Virulence Factor That Inhibits Tomato Immune Proteases
    Article Snippet: .. Efficient protein production using an adjusted bacterial expression system for secreted proteins To produce the selected proteins heterologously, we took advantage of the commercially available pFLAG-ATS expression system in Escherichia coli (Sigma-Aldrich). .. This expression system secretes N-terminally FLAG-tagged proteins into the growth medium using an N-terminal OmpA signal peptide to facilitate secretion.

    Ex Vivo:

    Article Title: Oleuropein Decreases Cyclooxygenase-2 and Interleukin-17 Expression and Attenuates Inflammatory Damage in Colonic Samples from Ulcerative Colitis Patients
    Article Snippet: Paragraph title: 2.2. Ex Vivo Organ Culture ... Then, the dishes were placed in an organ culture chamber at 37 °C in 95% O2 , 5% CO2 for 4 and 20 h, and biopsies were stimulated or not (untreated controls) with LPS from Escherichia coli (EC-LPS, Escherichia coli Serotype O127:B8 Lipopolysaccharide, Sigma-Aldrich, Milan, Italy) at a concentration of 1 μg/mL, to induce an inflammatory response, in the absence or presence of OLE at a final concentration of 3 mM.

    Western Blot:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ). .. For the Western blot analysis, cells were cultured in 1% HS (GE Healthcare) and 1% FBS (Thermo Fisher Scientific Inc.).

    Transformation Assay:

    Article Title: Novel fusion proteins for the antigen-specific staining and elimination of B cell receptor-positive cell populations demonstrated by a tetanus toxoid fragment C (TTC) model antigen
    Article Snippet: .. Expression of TTC-ETA’ and TTC in Escherichia coli and protein purification Escherichia coli BL21 (DE3) cells (Novagen, Darmstadt, Germany) were transformed with the TTC and TTC-ETA’ encoding expression vectors and the corresponding proteins were expressed into the periplasm under osmotic stress in the presence of compatible solutes [ ]. .. The protein was purified from the periplasmic fraction by immobilized metal-ion affinity chromatography (IMAC) using a Nickel-Sepharose (Ni-NTA) Superflow Cartridge (Qiagen, Hilden, Germany) on the ÄKTApurifier system (GE Healthcare Life Sciences, Freiburg, Germany) followed by a size-exclusion chromatography using a Superdex 200 (GE Healthcare).

    Over Expression:

    Article Title: Characterization of Aphanizomenon ovalisporum amidinotransferase involved in cylindrospermopsin synthesis
    Article Snippet: .. Two Escherichia coli strains were utilized: DH5α, for cloning and sequencing the aoa A gene into pET28b+ vector (Novagen, Madison, WI), and BL21 (DE3) used for protein overexpression. .. Cloning of aoaA and overexpression of AoaA Cyanobacteria DNA extraction and purification were carried out following the method developed by Smoker and Barnum , and modified by Neilan ( ).

    Electron Microscopy:

    Article Title: Heparin-binding protein is important for vascular leak in sepsis
    Article Snippet: .. Electron microscopy and histology preparations were compared to those obtained from mice sacrificed at 4 h after intraperitoneal injection of lipopolysaccharide (LPS) from Escherichia coli 0111:B4 (Sigma-Aldrich) in a dose of 0.25 mg [ ] after preparation as described above. .. Statistical analysis Comparisons between groups were made using the non-parametric Mann-Whitney test, Student’s t test, one-way ANOVA, and two-way repeated measures ANOVA as appropriate.

    Flow Cytometry:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: After primary cell culture (passages 1–2), the surface marker expression of hASCs was analyzed by flow cytometry (fluorescence-activated cell sorting) (FACSAria; Becton, Dickinson and Company, Erembodegem, Belgium, ) ( ) [ ]. .. BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: The purity of CD14+ monocytes and CD3+ T cell populations was usually higher than 90%, as evaluated by flow cytometry analysis (CyFlow Cube 6, Sysmex Partec GmbH, Görlitz, Germany) (data not shown). .. The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1.

    Chromatography:

    Article Title: Receptor tyrosine kinase‐like orphan receptor 1, a target of NKX2‐1/TTF‐1 lineage‐survival oncogene, inhibits apoptosis signal‐regulating kinase 1‐mediated pro‐apoptotic signaling in lung adenocarcinoma
    Article Snippet: Preparation of recombinant proteins The GST‐tagged ROR1 (intracellular domain) was expressed in Sf9 insect cells using a Gateway system (Invitrogen) and purified by glutathione‐affinity chromatography, as previously described. .. An Escherichia coli ‐expressing protein of MBP‐tagged kinase‐dead MKK6 was purchased from Millipore.

    Cell Culture:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: In qALP and Oil Red O analyses, Nunc 24-well plates (Thermo Fisher Scientific Inc.) and in mineralization studies CellBIND (Corning Inc., Corning, NY, ), 24-well plates were used. hASCs were cultured in 4 different culturing conditions: BM, BM supplemented with BMP-2, osteogenic medium (OM; supplemented with 10 mM β-glycerophosphate, 200 µM l -ascorbic acid 2-phosphate, and 5 nM dexamethasone), and OM supplemented with BMP-2. .. BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: The corneal fibroblasts were cultured overnight in 25 cm2 flasks, the medium with non‐attached cells was discarded and the incubation was continued in a new medium which was changed every 3 days, until 90% confluence was achieved. .. The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay).

    Article Title: Characterization of Aphanizomenon ovalisporum amidinotransferase involved in cylindrospermopsin synthesis
    Article Snippet: The strain was identified according to Komarek and Anagnostidis , and cultured in BG11 medium under continuous white light (50 μmol m-2 S-1 ) at 28°C. .. Two Escherichia coli strains were utilized: DH5α, for cloning and sequencing the aoa A gene into pET28b+ vector (Novagen, Madison, WI), and BL21 (DE3) used for protein overexpression.

    Sequencing:

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: .. The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay). .. For the dose‐response assay the PCCF were treated with different concentrations of LPS, PGN, and CpG‐ODN and incubated at the optimal time found in the time‐response assay.

    Article Title: Characterization of Aphanizomenon ovalisporum amidinotransferase involved in cylindrospermopsin synthesis
    Article Snippet: .. Two Escherichia coli strains were utilized: DH5α, for cloning and sequencing the aoa A gene into pET28b+ vector (Novagen, Madison, WI), and BL21 (DE3) used for protein overexpression. .. Cloning of aoaA and overexpression of AoaA Cyanobacteria DNA extraction and purification were carried out following the method developed by Smoker and Barnum , and modified by Neilan ( ).

    Injection:

    Article Title: Heparin-binding protein is important for vascular leak in sepsis
    Article Snippet: .. Electron microscopy and histology preparations were compared to those obtained from mice sacrificed at 4 h after intraperitoneal injection of lipopolysaccharide (LPS) from Escherichia coli 0111:B4 (Sigma-Aldrich) in a dose of 0.25 mg [ ] after preparation as described above. .. Statistical analysis Comparisons between groups were made using the non-parametric Mann-Whitney test, Student’s t test, one-way ANOVA, and two-way repeated measures ANOVA as appropriate.

    Recombinant:

    Article Title: Receptor tyrosine kinase‐like orphan receptor 1, a target of NKX2‐1/TTF‐1 lineage‐survival oncogene, inhibits apoptosis signal‐regulating kinase 1‐mediated pro‐apoptotic signaling in lung adenocarcinoma
    Article Snippet: Paragraph title: Preparation of recombinant proteins ... An Escherichia coli ‐expressing protein of MBP‐tagged kinase‐dead MKK6 was purchased from Millipore.

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: .. The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1. .. To determine the role of toll-like receptors (TLR)2 and TLR4 on the DCs status induced by ES L1, immature DCs were treated with blocking antibodies against TLR2 and/or TLR4 (10 µg/ml each, both from BioLegend), or isotype control antibody (anti-rat IgG, eBioscience), 1 h before the treatment with ES L1.

    Organ Culture:

    Article Title: Oleuropein Decreases Cyclooxygenase-2 and Interleukin-17 Expression and Attenuates Inflammatory Damage in Colonic Samples from Ulcerative Colitis Patients
    Article Snippet: .. Then, the dishes were placed in an organ culture chamber at 37 °C in 95% O2 , 5% CO2 for 4 and 20 h, and biopsies were stimulated or not (untreated controls) with LPS from Escherichia coli (EC-LPS, Escherichia coli Serotype O127:B8 Lipopolysaccharide, Sigma-Aldrich, Milan, Italy) at a concentration of 1 μg/mL, to induce an inflammatory response, in the absence or presence of OLE at a final concentration of 3 mM. ..

    Magnetic Cell Separation:

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: CD3+ T cells were isolated from PBMCs by negative MACS, using the Pan T cell isolation kit (Miltenyi Biotec), and they were used as responders in allogeneic coculture experiments with monocyte-derived DCs. .. The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1.

    Marker:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: After primary cell culture (passages 1–2), the surface marker expression of hASCs was analyzed by flow cytometry (fluorescence-activated cell sorting) (FACSAria; Becton, Dickinson and Company, Erembodegem, Belgium, ) ( ) [ ]. .. BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    In Vivo:

    Article Title: Targeting proteins for degradation
    Article Snippet: Bougdour A, Wickner S, Gottesman S. Modulating RssB activity: IraP, a novel regulator of sigma(S) stability in Escherichia coli. .. In vivo half-life of a protein is a function of its amino-terminal residue.

    Article Title: Heparin-binding protein is important for vascular leak in sepsis
    Article Snippet: Paragraph title: In vivo studies ... Electron microscopy and histology preparations were compared to those obtained from mice sacrificed at 4 h after intraperitoneal injection of lipopolysaccharide (LPS) from Escherichia coli 0111:B4 (Sigma-Aldrich) in a dose of 0.25 mg [ ] after preparation as described above.

    Fluorescence:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: After primary cell culture (passages 1–2), the surface marker expression of hASCs was analyzed by flow cytometry (fluorescence-activated cell sorting) (FACSAria; Becton, Dickinson and Company, Erembodegem, Belgium, ) ( ) [ ]. .. BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    Cell Stimulation:

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: Paragraph title: PCCF and cellular stimulation assay ... The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay).

    Isolation:

    Article Title: Characterization of Aphanizomenon ovalisporum amidinotransferase involved in cylindrospermopsin synthesis
    Article Snippet: Strains and vectors The strain A. ovalisporum UAM-MAO (Barón-Sola et al. ) was isolated from a Spanish pond of the Parque Juan Carlos I in Madrid (40° 27′ N; 3° 36′ W), used for various recreation activities. .. Two Escherichia coli strains were utilized: DH5α, for cloning and sequencing the aoa A gene into pET28b+ vector (Novagen, Madison, WI), and BL21 (DE3) used for protein overexpression.

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: CD3+ T cells were isolated from PBMCs by negative MACS, using the Pan T cell isolation kit (Miltenyi Biotec), and they were used as responders in allogeneic coculture experiments with monocyte-derived DCs. .. The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1.

    Size-exclusion Chromatography:

    Article Title: Novel fusion proteins for the antigen-specific staining and elimination of B cell receptor-positive cell populations demonstrated by a tetanus toxoid fragment C (TTC) model antigen
    Article Snippet: Expression of TTC-ETA’ and TTC in Escherichia coli and protein purification Escherichia coli BL21 (DE3) cells (Novagen, Darmstadt, Germany) were transformed with the TTC and TTC-ETA’ encoding expression vectors and the corresponding proteins were expressed into the periplasm under osmotic stress in the presence of compatible solutes [ ]. .. The protein was purified from the periplasmic fraction by immobilized metal-ion affinity chromatography (IMAC) using a Nickel-Sepharose (Ni-NTA) Superflow Cartridge (Qiagen, Hilden, Germany) on the ÄKTApurifier system (GE Healthcare Life Sciences, Freiburg, Germany) followed by a size-exclusion chromatography using a Superdex 200 (GE Healthcare).

    Labeling:

    Article Title: Brain Region-dependent Heterogeneity and Dose-dependent Difference in Transient Microglia Population Increase during Lipopolysaccharide-induced Inflammation
    Article Snippet: As the inflammatory stimulation, mice received a single intraperitoneal administration of LPS from Escherichia coli (serotype 055:B5 and 0111:B4; Sigma-Aldrich) and Salmonella enterica (serotype typhimurium; Sigma-Aldrich) at a dose of either 100 μg/kg or 1 mg/kg in pyrogen-free physiological saline (Otsuka Pharmaceutical Co. LTD., Tokushima, Japan). .. Regarding BrdU labeling, mice received BrdU (1 mg/ml) orally for 3 days through their drinking water to examine brain region-dependent heterogeneity and dose-dependent difference of LPS-induced microglia proliferation.

    Purification:

    Article Title: Elevated Carbon Monoxide to Carbon Dioxide Ratio in the Exhaled Breath of Mice Treated With a Single Dose of Lipopolysaccharide
    Article Snippet: .. Reagents Escherichia coli 0111:B4 LPS, which had been phenol-extracted and chromatographically purified by gel filtration and then passed through 0.2-µm pore-sized membrane as a 1.0-mg/mL aqueous solution, was from Sigma-Aldrich (catalog L5293). .. The LPS solution or water was further diluted for use in phosphate-buffered saline (pH 7.4) solution in endotoxin-free Milli-Q (Millipore) water.

    Article Title: Novel fusion proteins for the antigen-specific staining and elimination of B cell receptor-positive cell populations demonstrated by a tetanus toxoid fragment C (TTC) model antigen
    Article Snippet: Expression of TTC-ETA’ and TTC in Escherichia coli and protein purification Escherichia coli BL21 (DE3) cells (Novagen, Darmstadt, Germany) were transformed with the TTC and TTC-ETA’ encoding expression vectors and the corresponding proteins were expressed into the periplasm under osmotic stress in the presence of compatible solutes [ ]. .. The protein was purified from the periplasmic fraction by immobilized metal-ion affinity chromatography (IMAC) using a Nickel-Sepharose (Ni-NTA) Superflow Cartridge (Qiagen, Hilden, Germany) on the ÄKTApurifier system (GE Healthcare Life Sciences, Freiburg, Germany) followed by a size-exclusion chromatography using a Superdex 200 (GE Healthcare).

    Article Title: Receptor tyrosine kinase‐like orphan receptor 1, a target of NKX2‐1/TTF‐1 lineage‐survival oncogene, inhibits apoptosis signal‐regulating kinase 1‐mediated pro‐apoptotic signaling in lung adenocarcinoma
    Article Snippet: Preparation of recombinant proteins The GST‐tagged ROR1 (intracellular domain) was expressed in Sf9 insect cells using a Gateway system (Invitrogen) and purified by glutathione‐affinity chromatography, as previously described. .. An Escherichia coli ‐expressing protein of MBP‐tagged kinase‐dead MKK6 was purchased from Millipore.

    Protein Purification:

    Article Title: Novel fusion proteins for the antigen-specific staining and elimination of B cell receptor-positive cell populations demonstrated by a tetanus toxoid fragment C (TTC) model antigen
    Article Snippet: .. Expression of TTC-ETA’ and TTC in Escherichia coli and protein purification Escherichia coli BL21 (DE3) cells (Novagen, Darmstadt, Germany) were transformed with the TTC and TTC-ETA’ encoding expression vectors and the corresponding proteins were expressed into the periplasm under osmotic stress in the presence of compatible solutes [ ]. .. The protein was purified from the periplasmic fraction by immobilized metal-ion affinity chromatography (IMAC) using a Nickel-Sepharose (Ni-NTA) Superflow Cartridge (Qiagen, Hilden, Germany) on the ÄKTApurifier system (GE Healthcare Life Sciences, Freiburg, Germany) followed by a size-exclusion chromatography using a Superdex 200 (GE Healthcare).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay). .. The total RNA of treated and non‐treated cells was obtained with Trizol reagent (Invitrogen) for analysis, through RT‐PCR, mRNA expression of: α‐defensin‐3 (AD‐3), α‐defensin‐4 (AD‐4), α‐defensin‐5 (AD‐5), β‐defensin‐1 (BD‐1), β‐defensin‐2 (BD‐2), β‐defensin‐3 (BD‐3), cathelin related antimicrobial peptide (CRAMP), TLR‐1, TLR‐2, TLR‐4, TLR‐6, and TLR‐9.

    Protein Extraction:

    Article Title: Oleuropein Decreases Cyclooxygenase-2 and Interleukin-17 Expression and Attenuates Inflammatory Damage in Colonic Samples from Ulcerative Colitis Patients
    Article Snippet: Then, the dishes were placed in an organ culture chamber at 37 °C in 95% O2 , 5% CO2 for 4 and 20 h, and biopsies were stimulated or not (untreated controls) with LPS from Escherichia coli (EC-LPS, Escherichia coli Serotype O127:B8 Lipopolysaccharide, Sigma-Aldrich, Milan, Italy) at a concentration of 1 μg/mL, to induce an inflammatory response, in the absence or presence of OLE at a final concentration of 3 mM. .. One biopsy specimen from each treatment was collected after 4 h and embedded in paraffin for histologic evaluation, while the remaining samples were snap-frozen after 20 h and then stored at −80 °C for further protein extraction.

    Polyacrylamide Gel Electrophoresis:

    Article Title: Novel fusion proteins for the antigen-specific staining and elimination of B cell receptor-positive cell populations demonstrated by a tetanus toxoid fragment C (TTC) model antigen
    Article Snippet: Expression of TTC-ETA’ and TTC in Escherichia coli and protein purification Escherichia coli BL21 (DE3) cells (Novagen, Darmstadt, Germany) were transformed with the TTC and TTC-ETA’ encoding expression vectors and the corresponding proteins were expressed into the periplasm under osmotic stress in the presence of compatible solutes [ ]. .. The proteins were passed through a 0.22-μm sterile filter (Nalgene, Roskilde, Denmark) and analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) as previously described [ ].

    Activated Clotting Time Assay:

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: .. The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay). .. For the dose‐response assay the PCCF were treated with different concentrations of LPS, PGN, and CpG‐ODN and incubated at the optimal time found in the time‐response assay.

    Mouse Assay:

    Article Title: Brain Region-dependent Heterogeneity and Dose-dependent Difference in Transient Microglia Population Increase during Lipopolysaccharide-induced Inflammation
    Article Snippet: .. As the inflammatory stimulation, mice received a single intraperitoneal administration of LPS from Escherichia coli (serotype 055:B5 and 0111:B4; Sigma-Aldrich) and Salmonella enterica (serotype typhimurium; Sigma-Aldrich) at a dose of either 100 μg/kg or 1 mg/kg in pyrogen-free physiological saline (Otsuka Pharmaceutical Co. LTD., Tokushima, Japan). .. Regarding BrdU labeling, mice received BrdU (1 mg/ml) orally for 3 days through their drinking water to examine brain region-dependent heterogeneity and dose-dependent difference of LPS-induced microglia proliferation.

    Article Title: Heparin-binding protein is important for vascular leak in sepsis
    Article Snippet: .. Electron microscopy and histology preparations were compared to those obtained from mice sacrificed at 4 h after intraperitoneal injection of lipopolysaccharide (LPS) from Escherichia coli 0111:B4 (Sigma-Aldrich) in a dose of 0.25 mg [ ] after preparation as described above. .. Statistical analysis Comparisons between groups were made using the non-parametric Mann-Whitney test, Student’s t test, one-way ANOVA, and two-way repeated measures ANOVA as appropriate.

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: Briefly, Balb/c and NIH mice corneas were digested with 0.6U of dispase and 80U of collagenase solution (Invitrogen, Carlsbad, CA, USA) at 37°C for 1 hour, followed by 15 minutes of digestion with trypsin‐EDTA (Invitrogen, Carlsbad, CA, USA) at 37°C. .. The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay).

    SDS Page:

    Article Title: Novel fusion proteins for the antigen-specific staining and elimination of B cell receptor-positive cell populations demonstrated by a tetanus toxoid fragment C (TTC) model antigen
    Article Snippet: Expression of TTC-ETA’ and TTC in Escherichia coli and protein purification Escherichia coli BL21 (DE3) cells (Novagen, Darmstadt, Germany) were transformed with the TTC and TTC-ETA’ encoding expression vectors and the corresponding proteins were expressed into the periplasm under osmotic stress in the presence of compatible solutes [ ]. .. The proteins were passed through a 0.22-μm sterile filter (Nalgene, Roskilde, Denmark) and analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) as previously described [ ].

    Plasmid Preparation:

    Article Title: Characterization of Aphanizomenon ovalisporum amidinotransferase involved in cylindrospermopsin synthesis
    Article Snippet: .. Two Escherichia coli strains were utilized: DH5α, for cloning and sequencing the aoa A gene into pET28b+ vector (Novagen, Madison, WI), and BL21 (DE3) used for protein overexpression. .. Cloning of aoaA and overexpression of AoaA Cyanobacteria DNA extraction and purification were carried out following the method developed by Smoker and Barnum , and modified by Neilan ( ).

    Functional Assay:

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: .. The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1. .. To determine the role of toll-like receptors (TLR)2 and TLR4 on the DCs status induced by ES L1, immature DCs were treated with blocking antibodies against TLR2 and/or TLR4 (10 µg/ml each, both from BioLegend), or isotype control antibody (anti-rat IgG, eBioscience), 1 h before the treatment with ES L1.

    Affinity Chromatography:

    Article Title: Novel fusion proteins for the antigen-specific staining and elimination of B cell receptor-positive cell populations demonstrated by a tetanus toxoid fragment C (TTC) model antigen
    Article Snippet: Expression of TTC-ETA’ and TTC in Escherichia coli and protein purification Escherichia coli BL21 (DE3) cells (Novagen, Darmstadt, Germany) were transformed with the TTC and TTC-ETA’ encoding expression vectors and the corresponding proteins were expressed into the periplasm under osmotic stress in the presence of compatible solutes [ ]. .. The protein was purified from the periplasmic fraction by immobilized metal-ion affinity chromatography (IMAC) using a Nickel-Sepharose (Ni-NTA) Superflow Cartridge (Qiagen, Hilden, Germany) on the ÄKTApurifier system (GE Healthcare Life Sciences, Freiburg, Germany) followed by a size-exclusion chromatography using a Superdex 200 (GE Healthcare).

    Centrifugation:

    Article Title: Screen of Non-annotated Small Secreted Proteins of Pseudomonas syringae Reveals a Virulence Factor That Inhibits Tomato Immune Proteases
    Article Snippet: Efficient protein production using an adjusted bacterial expression system for secreted proteins To produce the selected proteins heterologously, we took advantage of the commercially available pFLAG-ATS expression system in Escherichia coli (Sigma-Aldrich). .. The growth medium of E . coli cultures has relatively low protein content and is easy to collect as supernatant after centrifugation, making this expression system ideal to produce small secreted bacterial proteins.

    Concentration Assay:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ). .. BMP-2 was used in concentration of 100 ng/ml unless otherwise mentioned.

    Article Title: Oleuropein Decreases Cyclooxygenase-2 and Interleukin-17 Expression and Attenuates Inflammatory Damage in Colonic Samples from Ulcerative Colitis Patients
    Article Snippet: .. Then, the dishes were placed in an organ culture chamber at 37 °C in 95% O2 , 5% CO2 for 4 and 20 h, and biopsies were stimulated or not (untreated controls) with LPS from Escherichia coli (EC-LPS, Escherichia coli Serotype O127:B8 Lipopolysaccharide, Sigma-Aldrich, Milan, Italy) at a concentration of 1 μg/mL, to induce an inflammatory response, in the absence or presence of OLE at a final concentration of 3 mM. ..

    CTG Assay:

    Article Title: Expression of CRAMP via PGN-TLR-2 and of ?-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts
    Article Snippet: .. The Balb/c PCCF were treated with 10 μg/ml of LPS from Escherichia coli 0111:B4 (Sigma chemical, St Louis, MO, USA), 10 μg/ml of PGN from Staphylococcus aureus (Sigma Fluka, St Louis, MO, USA), 3 ng/ml of CpG‐ODN (TCC ATG ACG TTC CTG ACG TT; Invitrogen, Carlsbad CA, USA), or with 3 ng/ml of irrelevant oligonucleotide without CpG sequence (TCC AGG ACT TCT CTT CAG GTT; Invitrogen) according to the stimulus, then dissolved in fresh MEM medium for different times: 0, 3, 6, 12, 24, 48, and 72 hours (time‐response assay). .. For the dose‐response assay the PCCF were treated with different concentrations of LPS, PGN, and CpG‐ODN and incubated at the optimal time found in the time‐response assay.

    FACS:

    Article Title: Bone Morphogenetic Protein-2 Induces Donor-Dependent Osteogenic and Adipogenic Differentiation in Human Adipose Stem Cells
    Article Snippet: After primary cell culture (passages 1–2), the surface marker expression of hASCs was analyzed by flow cytometry (fluorescence-activated cell sorting) (FACSAria; Becton, Dickinson and Company, Erembodegem, Belgium, ) ( ) [ ]. .. BMP-2 originated from either mammalian cultures (Chinese hamster ovary cells [CHO]; R & D Systems, Minneapolis, MN, ) or from Escherichia coli (Sigma-Aldrich, St. Louis, MO, ).

    Article Title: Trichinella spiralis Excretory–Secretory Products Induce Tolerogenic Properties in Human Dendritic Cells via Toll-Like Receptors 2 and 4
    Article Snippet: The monocytes were isolated by negative magnetic-activated cell sorting (MACS) from PBMCs, using the Monocyte Isolation kit (Myltenil Biotec), following their cultivation in CellGro DC medium (Cell Genix), supplemented with 100 ng/ml of human recombinant GM-CSF and 20 ng/ml of human recombinant IL-4 (both from R & D Systems) (complete DC growth medium). .. The impact of ES L1 on the maturation of DCs was determined by adding ES L1 antigens to the culture of immature DCs on Day 4, for 48 h. To induce mature DCs, the cells were stimulated with LPS from Escherichia coli (500 ng/ml, Sigma-Aldrich) and human recombinant IFN-γ (50 ng/ml, R & D Systems) on Day 5, for the next 24 h. In some experiments, LPS (500 ng/ml) or polyinosininc:polycytidylic acid [Poly (I:C), 10 µg/ml] were used instead of LPS/IFN-γ, to investigate the stability of phenotypical and functional characteristics of DCs acquired after the pulsing with ES L1.

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    Millipore escherichia coli novablue competent cells
    Escherichia Coli Novablue Competent Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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