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Millipore protease
Protease, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 513 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/protease/product/Millipore
Average 99 stars, based on 513 article reviews
Price from $9.99 to $1999.99
protease - by Bioz Stars, 2020-07
99/100 stars

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Protease Inhibitor:

Article Title: The serine protease hepsin mediates urinary secretion and polymerisation of Zona Pellucida domain protein uromodulin
Article Snippet: .. After 24 h, the medium was replaced with Optimem supplemented with 0.1% protease inhibitor cocktail (PIC) (P8340, Sigma-Aldrich, Saint Louis, MO), 0.1% DMSO, 0.1 mM AEBSF, 0.08 μM aprotinin, 1.4 μM E64, 4 μM bestatin, 2 μM leupeptin and 1.5 μM pepstatin A (Sigma-Aldrich). .. Immunofluorescence analysis was performed after 24 hr of treatment.

Article Title: Membrane Cholesterol Modulates LOX-1 Shedding in Endothelial Cells
Article Snippet: .. Phenylmethylsulfonylfuoride (PMSF) (Euroclone, Devon, UK), pepstatin A (Boehringer, Mannheim, DE), GM6001 inhibitor (Biomol Research Lab Inc.), protease inhibitor cocktail set III (AEBSF 100mM; aprotinin 80μM; bestatin 5mM; E-64 1,5mM; leupeptin 2mM; pepstatin A 1mM) (Calbiochem, La Jolla, CA) were used for specific protease class inhibition experiments. .. Ox-LDL preparation, labelling and fluorescence analysis Human LDL was prepared from fresh healthy normolipidemic plasma of volunteers by ultracentrifugation [ ] and in vitro oxidized as described (Matarazzo et al., 2012).

Article Title: Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase
Article Snippet: .. Co-immunoprecipitation Uninfected or E. chaffeensis -infected THP-1 cells at 2 d p.i. were lysed in modified lysis buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP40 [USB, 19638], 5% glycerol, and 1% protease inhibitor cocktail III [Calbiochem, 539134]) for 15 min and immunoprecipitated for 2 h with rabbit anti-Etf-1, -BECN1, or -PIK3C3, or mouse anti-RAB5 IgG that was cross-linked to protein A/G-magnetic beads (Pierce, 88805). .. Normal rabbit (Santa Cruz Biotechnology, sc-2027) or mouse IgG (Santa Cruz Biotechnology, sc-2025) was used as a negative control for immunoprecipitation.

Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace
Article Snippet: .. For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C. .. Protein lysate samples were used for mouse neutrophil elastase/ELA2 and MPO ELISA (R & D Systems, MN) per manufacturers protocol.

Article Title: Nasal Immunity to Staphylococcal Toxic Shock Is Controlled by the Nasopharynx-Associated Lymphoid Tissue ▿Nasal Immunity to Staphylococcal Toxic Shock Is Controlled by the Nasopharynx-Associated Lymphoid Tissue ▿ †Nasal Immunity to Staphylococcal Toxic Shock Is Controlled by the Nasopharynx-Associated Lymphoid Tissue ▿ † ‡
Article Snippet: .. The collected rinse (5 to 8 μl) was deposited in a fresh centrifuge tube containing 10 μl PBS with 2× protease inhibitor cocktail set III (Calbiochem, San Diego, CA). .. Nasal lavage was collected from euthanized (CO2 asphyxiation, followed by cervical dislocation) mice by carefully introducing 20 to 30 μl PBS with a pipette through one nostril.

Article Title: Src family kinases Fyn and Lyn are constitutively activated and mediate plasmacytoid dendritic cell responses
Article Snippet: .. Immunoprecipitation and immunoblotting CAL-1 cells were pre-treated with either PP2 or DMSO control for 1 h, stimulated with R848 (1 μg ml−1 ) and subsequently lysed with immunoprecipitation lysis buffer (Thermo Scientific, Rockford, IL, USA) supplemented with protease inhibitor cocktail set III (EMD Millipore) and phosphatase inhibitor cocktail set I and II (EMD Millipore) for immunoprecipitation analysis, with RIPA buffer (Thermo Scientific) supplemented with protease inhibitor cocktail set III (EMD Millipore) for immunoblotting analysis or processed with the Subcellular Protein Fractionation Kit for Cultured Cells (Thermo Scientific) for subcellular fractionation. ..

Article Title: The C-Terminal Extension Unique to the Long Isoform of the Shelterin Component TIN2 Enhances Its Interaction with TRF2 in a Phosphorylation- and Dyskeratosis Congenita Cluster-Dependent Fashion
Article Snippet: .. Cells were resuspended in ice-cold lysis buffer (50 mM Tris-HCl at pH 7.5, 1 mM EDTA, 400 mM NaCl, 1% Triton X-100, 0.1% SDS, 1 mM dithiothreitol [DTT], 1 mM phenylmethylsulfonyl fluoride [PMSF], and 1× protease inhibitor cocktail III [Calbiochem]) and incubated for 10 min on ice prior to addition of an equal amount of ice-cold water. .. Protein concentration was determined using the bicinchoninic acid (BCA) protein assay kit (Pierce).

Infection:

Article Title: Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase
Article Snippet: .. Co-immunoprecipitation Uninfected or E. chaffeensis -infected THP-1 cells at 2 d p.i. were lysed in modified lysis buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP40 [USB, 19638], 5% glycerol, and 1% protease inhibitor cocktail III [Calbiochem, 539134]) for 15 min and immunoprecipitated for 2 h with rabbit anti-Etf-1, -BECN1, or -PIK3C3, or mouse anti-RAB5 IgG that was cross-linked to protein A/G-magnetic beads (Pierce, 88805). .. Normal rabbit (Santa Cruz Biotechnology, sc-2027) or mouse IgG (Santa Cruz Biotechnology, sc-2025) was used as a negative control for immunoprecipitation.

Immunoprecipitation:

Article Title: Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase
Article Snippet: .. Co-immunoprecipitation Uninfected or E. chaffeensis -infected THP-1 cells at 2 d p.i. were lysed in modified lysis buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP40 [USB, 19638], 5% glycerol, and 1% protease inhibitor cocktail III [Calbiochem, 539134]) for 15 min and immunoprecipitated for 2 h with rabbit anti-Etf-1, -BECN1, or -PIK3C3, or mouse anti-RAB5 IgG that was cross-linked to protein A/G-magnetic beads (Pierce, 88805). .. Normal rabbit (Santa Cruz Biotechnology, sc-2027) or mouse IgG (Santa Cruz Biotechnology, sc-2025) was used as a negative control for immunoprecipitation.

Article Title: Src family kinases Fyn and Lyn are constitutively activated and mediate plasmacytoid dendritic cell responses
Article Snippet: .. Immunoprecipitation and immunoblotting CAL-1 cells were pre-treated with either PP2 or DMSO control for 1 h, stimulated with R848 (1 μg ml−1 ) and subsequently lysed with immunoprecipitation lysis buffer (Thermo Scientific, Rockford, IL, USA) supplemented with protease inhibitor cocktail set III (EMD Millipore) and phosphatase inhibitor cocktail set I and II (EMD Millipore) for immunoprecipitation analysis, with RIPA buffer (Thermo Scientific) supplemented with protease inhibitor cocktail set III (EMD Millipore) for immunoblotting analysis or processed with the Subcellular Protein Fractionation Kit for Cultured Cells (Thermo Scientific) for subcellular fractionation. ..

Incubation:

Article Title: The C-Terminal Extension Unique to the Long Isoform of the Shelterin Component TIN2 Enhances Its Interaction with TRF2 in a Phosphorylation- and Dyskeratosis Congenita Cluster-Dependent Fashion
Article Snippet: .. Cells were resuspended in ice-cold lysis buffer (50 mM Tris-HCl at pH 7.5, 1 mM EDTA, 400 mM NaCl, 1% Triton X-100, 0.1% SDS, 1 mM dithiothreitol [DTT], 1 mM phenylmethylsulfonyl fluoride [PMSF], and 1× protease inhibitor cocktail III [Calbiochem]) and incubated for 10 min on ice prior to addition of an equal amount of ice-cold water. .. Protein concentration was determined using the bicinchoninic acid (BCA) protein assay kit (Pierce).

Inhibition:

Article Title: Membrane Cholesterol Modulates LOX-1 Shedding in Endothelial Cells
Article Snippet: .. Phenylmethylsulfonylfuoride (PMSF) (Euroclone, Devon, UK), pepstatin A (Boehringer, Mannheim, DE), GM6001 inhibitor (Biomol Research Lab Inc.), protease inhibitor cocktail set III (AEBSF 100mM; aprotinin 80μM; bestatin 5mM; E-64 1,5mM; leupeptin 2mM; pepstatin A 1mM) (Calbiochem, La Jolla, CA) were used for specific protease class inhibition experiments. .. Ox-LDL preparation, labelling and fluorescence analysis Human LDL was prepared from fresh healthy normolipidemic plasma of volunteers by ultracentrifugation [ ] and in vitro oxidized as described (Matarazzo et al., 2012).

Fractionation:

Article Title: Src family kinases Fyn and Lyn are constitutively activated and mediate plasmacytoid dendritic cell responses
Article Snippet: .. Immunoprecipitation and immunoblotting CAL-1 cells were pre-treated with either PP2 or DMSO control for 1 h, stimulated with R848 (1 μg ml−1 ) and subsequently lysed with immunoprecipitation lysis buffer (Thermo Scientific, Rockford, IL, USA) supplemented with protease inhibitor cocktail set III (EMD Millipore) and phosphatase inhibitor cocktail set I and II (EMD Millipore) for immunoprecipitation analysis, with RIPA buffer (Thermo Scientific) supplemented with protease inhibitor cocktail set III (EMD Millipore) for immunoblotting analysis or processed with the Subcellular Protein Fractionation Kit for Cultured Cells (Thermo Scientific) for subcellular fractionation. ..

Cell Culture:

Article Title: Src family kinases Fyn and Lyn are constitutively activated and mediate plasmacytoid dendritic cell responses
Article Snippet: .. Immunoprecipitation and immunoblotting CAL-1 cells were pre-treated with either PP2 or DMSO control for 1 h, stimulated with R848 (1 μg ml−1 ) and subsequently lysed with immunoprecipitation lysis buffer (Thermo Scientific, Rockford, IL, USA) supplemented with protease inhibitor cocktail set III (EMD Millipore) and phosphatase inhibitor cocktail set I and II (EMD Millipore) for immunoprecipitation analysis, with RIPA buffer (Thermo Scientific) supplemented with protease inhibitor cocktail set III (EMD Millipore) for immunoblotting analysis or processed with the Subcellular Protein Fractionation Kit for Cultured Cells (Thermo Scientific) for subcellular fractionation. ..

Modification:

Article Title: Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase
Article Snippet: .. Co-immunoprecipitation Uninfected or E. chaffeensis -infected THP-1 cells at 2 d p.i. were lysed in modified lysis buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP40 [USB, 19638], 5% glycerol, and 1% protease inhibitor cocktail III [Calbiochem, 539134]) for 15 min and immunoprecipitated for 2 h with rabbit anti-Etf-1, -BECN1, or -PIK3C3, or mouse anti-RAB5 IgG that was cross-linked to protein A/G-magnetic beads (Pierce, 88805). .. Normal rabbit (Santa Cruz Biotechnology, sc-2027) or mouse IgG (Santa Cruz Biotechnology, sc-2025) was used as a negative control for immunoprecipitation.

Lysis:

Article Title: Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase
Article Snippet: .. Co-immunoprecipitation Uninfected or E. chaffeensis -infected THP-1 cells at 2 d p.i. were lysed in modified lysis buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP40 [USB, 19638], 5% glycerol, and 1% protease inhibitor cocktail III [Calbiochem, 539134]) for 15 min and immunoprecipitated for 2 h with rabbit anti-Etf-1, -BECN1, or -PIK3C3, or mouse anti-RAB5 IgG that was cross-linked to protein A/G-magnetic beads (Pierce, 88805). .. Normal rabbit (Santa Cruz Biotechnology, sc-2027) or mouse IgG (Santa Cruz Biotechnology, sc-2025) was used as a negative control for immunoprecipitation.

Article Title: Src family kinases Fyn and Lyn are constitutively activated and mediate plasmacytoid dendritic cell responses
Article Snippet: .. Immunoprecipitation and immunoblotting CAL-1 cells were pre-treated with either PP2 or DMSO control for 1 h, stimulated with R848 (1 μg ml−1 ) and subsequently lysed with immunoprecipitation lysis buffer (Thermo Scientific, Rockford, IL, USA) supplemented with protease inhibitor cocktail set III (EMD Millipore) and phosphatase inhibitor cocktail set I and II (EMD Millipore) for immunoprecipitation analysis, with RIPA buffer (Thermo Scientific) supplemented with protease inhibitor cocktail set III (EMD Millipore) for immunoblotting analysis or processed with the Subcellular Protein Fractionation Kit for Cultured Cells (Thermo Scientific) for subcellular fractionation. ..

Article Title: The C-Terminal Extension Unique to the Long Isoform of the Shelterin Component TIN2 Enhances Its Interaction with TRF2 in a Phosphorylation- and Dyskeratosis Congenita Cluster-Dependent Fashion
Article Snippet: .. Cells were resuspended in ice-cold lysis buffer (50 mM Tris-HCl at pH 7.5, 1 mM EDTA, 400 mM NaCl, 1% Triton X-100, 0.1% SDS, 1 mM dithiothreitol [DTT], 1 mM phenylmethylsulfonyl fluoride [PMSF], and 1× protease inhibitor cocktail III [Calbiochem]) and incubated for 10 min on ice prior to addition of an equal amount of ice-cold water. .. Protein concentration was determined using the bicinchoninic acid (BCA) protein assay kit (Pierce).

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  • 99
    Millipore protease inhibitor cocktail set iii
    Coinfection with H. polygyrus increases neutrophil transepithelial migration into the P. aeruginosa infected airspace of the lung. FACS analysis using Ly6G and CD11b markers to isolate the neutrophil population in the ( a ) <t>BAL,</t> ( b ) lung tissue and ( c ) peripheral blood (PB), data are shown as mean ± SD, and are representative of <t>three</t> independent experiments. * P
    Protease Inhibitor Cocktail Set Iii, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 616 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protease inhibitor cocktail set iii/product/Millipore
    Average 99 stars, based on 616 article reviews
    Price from $9.99 to $1999.99
    protease inhibitor cocktail set iii - by Bioz Stars, 2020-07
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    88
    Millipore furin like protease
    Characterization of the MACIT protein <t>COL-99</t> of C. elegans . a Schematic structure of the COL-99 and COL-99::EGFP::FLAG polypeptides. The domain structure is presented as in Fig. 1 and cysteines ( c ) are also indicated. The regions used as antigens for the antibodies AB5625.11 and AB693 are marked. The C-terminal tag of COL-99::EGFP::FLAG is shown as a green circle. b Western blot analysis of subcellular localization of recombinant COL-99 in CHO cells. From the left to right: control lysate, total lysate of CHO cells expressing recombinant COL-99, concentrated medium sample, in the upper and lower panels detected with antibody AB5625.11 ( upper ) or AB693 ( lower ); and additional PBS-soluble proteins, and Triton X-100-extracted protein fraction from CHO cells expressing COL-99 in the lower panel only detected with AB693. c Immunofluorescent staining of CHO cells expressing COL-99. The signal was detected by antibody AB5625.11 and secondary anti-rabbit IgG conjugated with AlexaFluor 594. d Control staining with the secondary antibody only. Bars, 20 μm. e Inhibition of COL-99 ectodomain shedding by <t>Furin</t> Inhibitor I. Right lane, COL-99 expressing CHO cell cultures treated with 2 μM Furin Inhibitor I. Left lane, cells treated with an equivalent volume of methanol as solvent control. β-tubulin was used as a loading control in ( b ) and ( e )
    Furin Like Protease, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/furin like protease/product/Millipore
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    furin like protease - by Bioz Stars, 2020-07
    88/100 stars
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    85
    Millipore tnf α protease inhibitor tapi 1
    ErbB4 transactivation by <t>TNF</t> promotes increased Akt activation. YAMC-vector and YAMC-ErbB4 cells were incubated with 10 μM <t>TAPI-1</t> for 30 min ( A ) or with ErbB4 blocking antibody for 1 h ( B ) before treatment with 100 ng/ml TNF for 15 min. Cell lysates
    Tnf α Protease Inhibitor Tapi 1, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tnf α protease inhibitor tapi 1/product/Millipore
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tnf α protease inhibitor tapi 1 - by Bioz Stars, 2020-07
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    Coinfection with H. polygyrus increases neutrophil transepithelial migration into the P. aeruginosa infected airspace of the lung. FACS analysis using Ly6G and CD11b markers to isolate the neutrophil population in the ( a ) BAL, ( b ) lung tissue and ( c ) peripheral blood (PB), data are shown as mean ± SD, and are representative of three independent experiments. * P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Coinfection with H. polygyrus increases neutrophil transepithelial migration into the P. aeruginosa infected airspace of the lung. FACS analysis using Ly6G and CD11b markers to isolate the neutrophil population in the ( a ) BAL, ( b ) lung tissue and ( c ) peripheral blood (PB), data are shown as mean ± SD, and are representative of three independent experiments. * P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: Migration, Infection, FACS

    Helminth-induced increase in neutrophil migration during P. aeruginosa infection is correlated with increased MPO and ELA2 levels within the airspace. Protein extracts were prepared from BAL samples and analyzed for ( a ) neutrophil myeloperoxidase (MPO) and ( b ) elastase (ELA2). ( c ) LDH levels were assessed in the cell-free supernatant of the BAL to assess cytotoxicity. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Helminth-induced increase in neutrophil migration during P. aeruginosa infection is correlated with increased MPO and ELA2 levels within the airspace. Protein extracts were prepared from BAL samples and analyzed for ( a ) neutrophil myeloperoxidase (MPO) and ( b ) elastase (ELA2). ( c ) LDH levels were assessed in the cell-free supernatant of the BAL to assess cytotoxicity. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: Migration, Infection

    Coinfection with H. polygyrus does not impact macrophage levels within the airspace, lung tissue or circulation during P. aeruginosa infection of the airspace. FACS analysis using F4/80 and CD11b markers to isolate the macrophage populations in the ( a ) BAL, ( b ) lung tissue and ( c ) monocytes in peripheral blood. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Coinfection with H. polygyrus does not impact macrophage levels within the airspace, lung tissue or circulation during P. aeruginosa infection of the airspace. FACS analysis using F4/80 and CD11b markers to isolate the macrophage populations in the ( a ) BAL, ( b ) lung tissue and ( c ) monocytes in peripheral blood. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: Infection, FACS

    Coinfection with H. polygyrus increases cellular transepithelial migration into the lung airspace. After 2 weeks of helminth infection, the mice were infected with/without P. aeruginosa through intranasal inoculation. Mice were placed in the cage for 6 hours, and after incubation mice were sacrificed, BAL and lung tissue were obtained. ( a ) The result of BAL cell count. Data are shown as mean ± SD and are representative of three independent experiments. ** P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Coinfection with H. polygyrus increases cellular transepithelial migration into the lung airspace. After 2 weeks of helminth infection, the mice were infected with/without P. aeruginosa through intranasal inoculation. Mice were placed in the cage for 6 hours, and after incubation mice were sacrificed, BAL and lung tissue were obtained. ( a ) The result of BAL cell count. Data are shown as mean ± SD and are representative of three independent experiments. ** P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: Migration, Infection, Mouse Assay, Incubation, Cell Counting

    Coinfection with H. polygyrus during P. aeruginosa acute pneumonia increases the CD4 + T-cells population in the airspace and lung tissues. FACS analysis using CD4 marker to isolate the CD4 + T-cell population in the ( a ) BAL, ( b ) lung tissue and ( c ) peripheral blood. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Coinfection with H. polygyrus during P. aeruginosa acute pneumonia increases the CD4 + T-cells population in the airspace and lung tissues. FACS analysis using CD4 marker to isolate the CD4 + T-cell population in the ( a ) BAL, ( b ) lung tissue and ( c ) peripheral blood. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: FACS, Marker

    The protein complementation assay confirms the effects of the R282H mutation and a phosphorylation site mutation on TIN2L interaction with TRF2. (A) Quantification of fluorescence from coexpression of V1-TRF2 with TIN2L-V2, TIN2L-D391K+D395K-V2, or TIN2L-R282H-V2. Error bars represent the SDs from three separate transfections each measured in triplicate. *, P

    Journal: Molecular and Cellular Biology

    Article Title: The C-Terminal Extension Unique to the Long Isoform of the Shelterin Component TIN2 Enhances Its Interaction with TRF2 in a Phosphorylation- and Dyskeratosis Congenita Cluster-Dependent Fashion

    doi: 10.1128/MCB.00025-18

    Figure Lengend Snippet: The protein complementation assay confirms the effects of the R282H mutation and a phosphorylation site mutation on TIN2L interaction with TRF2. (A) Quantification of fluorescence from coexpression of V1-TRF2 with TIN2L-V2, TIN2L-D391K+D395K-V2, or TIN2L-R282H-V2. Error bars represent the SDs from three separate transfections each measured in triplicate. *, P

    Article Snippet: Cells were resuspended in ice-cold lysis buffer (50 mM Tris-HCl at pH 7.5, 1 mM EDTA, 400 mM NaCl, 1% Triton X-100, 0.1% SDS, 1 mM dithiothreitol [DTT], 1 mM phenylmethylsulfonyl fluoride [PMSF], and 1× protease inhibitor cocktail III [Calbiochem]) and incubated for 10 min on ice prior to addition of an equal amount of ice-cold water.

    Techniques: Mutagenesis, Fluorescence, Transfection

    Characterization of the MACIT protein COL-99 of C. elegans . a Schematic structure of the COL-99 and COL-99::EGFP::FLAG polypeptides. The domain structure is presented as in Fig. 1 and cysteines ( c ) are also indicated. The regions used as antigens for the antibodies AB5625.11 and AB693 are marked. The C-terminal tag of COL-99::EGFP::FLAG is shown as a green circle. b Western blot analysis of subcellular localization of recombinant COL-99 in CHO cells. From the left to right: control lysate, total lysate of CHO cells expressing recombinant COL-99, concentrated medium sample, in the upper and lower panels detected with antibody AB5625.11 ( upper ) or AB693 ( lower ); and additional PBS-soluble proteins, and Triton X-100-extracted protein fraction from CHO cells expressing COL-99 in the lower panel only detected with AB693. c Immunofluorescent staining of CHO cells expressing COL-99. The signal was detected by antibody AB5625.11 and secondary anti-rabbit IgG conjugated with AlexaFluor 594. d Control staining with the secondary antibody only. Bars, 20 μm. e Inhibition of COL-99 ectodomain shedding by Furin Inhibitor I. Right lane, COL-99 expressing CHO cell cultures treated with 2 μM Furin Inhibitor I. Left lane, cells treated with an equivalent volume of methanol as solvent control. β-tubulin was used as a loading control in ( b ) and ( e )

    Journal: BMC Evolutionary Biology

    Article Title: Membrane-associated collagens with interrupted triple-helices (MACITs): evolution from a bilaterian common ancestor and functional conservation in C. elegans

    doi: 10.1186/s12862-015-0554-3

    Figure Lengend Snippet: Characterization of the MACIT protein COL-99 of C. elegans . a Schematic structure of the COL-99 and COL-99::EGFP::FLAG polypeptides. The domain structure is presented as in Fig. 1 and cysteines ( c ) are also indicated. The regions used as antigens for the antibodies AB5625.11 and AB693 are marked. The C-terminal tag of COL-99::EGFP::FLAG is shown as a green circle. b Western blot analysis of subcellular localization of recombinant COL-99 in CHO cells. From the left to right: control lysate, total lysate of CHO cells expressing recombinant COL-99, concentrated medium sample, in the upper and lower panels detected with antibody AB5625.11 ( upper ) or AB693 ( lower ); and additional PBS-soluble proteins, and Triton X-100-extracted protein fraction from CHO cells expressing COL-99 in the lower panel only detected with AB693. c Immunofluorescent staining of CHO cells expressing COL-99. The signal was detected by antibody AB5625.11 and secondary anti-rabbit IgG conjugated with AlexaFluor 594. d Control staining with the secondary antibody only. Bars, 20 μm. e Inhibition of COL-99 ectodomain shedding by Furin Inhibitor I. Right lane, COL-99 expressing CHO cell cultures treated with 2 μM Furin Inhibitor I. Left lane, cells treated with an equivalent volume of methanol as solvent control. β-tubulin was used as a loading control in ( b ) and ( e )

    Article Snippet: To test the shedding of the COL-99 ectodomain by furin-like protease, Furin Inhibitor I (Calbiochem) in 10 μl methanol at final concentrations of 0, 1, 2, 5, or 10 μM was added daily to CHO/COL-99 cells at ~80 % confluency growing in a 12-well plate with 1 ml of serum-free culture medium per well.

    Techniques: Western Blot, Recombinant, Expressing, Staining, Inhibition

    ErbB4 transactivation by TNF promotes increased Akt activation. YAMC-vector and YAMC-ErbB4 cells were incubated with 10 μM TAPI-1 for 30 min ( A ) or with ErbB4 blocking antibody for 1 h ( B ) before treatment with 100 ng/ml TNF for 15 min. Cell lysates

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: TNF-? converting enzyme-mediated ErbB4 transactivation by TNF promotes colonic epithelial cell survival

    doi: 10.1152/ajpgi.00057.2011

    Figure Lengend Snippet: ErbB4 transactivation by TNF promotes increased Akt activation. YAMC-vector and YAMC-ErbB4 cells were incubated with 10 μM TAPI-1 for 30 min ( A ) or with ErbB4 blocking antibody for 1 h ( B ) before treatment with 100 ng/ml TNF for 15 min. Cell lysates

    Article Snippet: TNF-α protease inhibitor (TAPI)-1 and GM6001 were purchased from EMD Chemicals (Darmstadt, Germany).

    Techniques: Activation Assay, Plasmid Preparation, Incubation, Blocking Assay

    ErbB4 transactivation by TNF protects colonic epithelial cells from cytokine-induced apoptosis. YAMC cells expressing either vector (YAMC-vector) or ErbB4 (YAMC-ErbB4) were incubated with 10 μM TAPI-1 for 30 min ( A ) or ErbB4 blocking antibody

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: TNF-? converting enzyme-mediated ErbB4 transactivation by TNF promotes colonic epithelial cell survival

    doi: 10.1152/ajpgi.00057.2011

    Figure Lengend Snippet: ErbB4 transactivation by TNF protects colonic epithelial cells from cytokine-induced apoptosis. YAMC cells expressing either vector (YAMC-vector) or ErbB4 (YAMC-ErbB4) were incubated with 10 μM TAPI-1 for 30 min ( A ) or ErbB4 blocking antibody

    Article Snippet: TNF-α protease inhibitor (TAPI)-1 and GM6001 were purchased from EMD Chemicals (Darmstadt, Germany).

    Techniques: Expressing, Plasmid Preparation, Incubation, Blocking Assay