p22 phox ab75941  (Thermo Fisher)


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    Thermo Fisher p22 phox ab75941
    Confocal imaging of living cells (Left A – F) ( A , B ): monocytes isolated from blood of healthy donors; the red signal is the NOX2 and mitotracker in ( A , B ), respectively, the grey signal corresponds to the mitotracker and CellRox deep red reagent in ( A , B ), respectively; ( C , D ) Monocyte-derived macrophages (with CSF1-without NS1 treatment) using the same markers used for the living monocytes in ( A , B ); ( E , F ) the macrophages were treated with NS1 after their CSF1-induced differentiation from monocytes using the same markers as in ( A , B ); the green signal in ( F ) is the fluorescence of NS1; the ROS levels are quantified by the cell-permeable CellROX deep red reagent shown in light grey, the mitochondria are monitored by the red mitotracker reagent as quantified in ( G ); Co-localization experiments in fixed cells (Right H – L) The images ( H – L ) present co-localization experiments in fixed monocytes or CSF1-treated monocytes differentiated into macrophages . Each antibody is mentioned above each image, the merge, representing the colocalization is shown in yellow. The intensity of the colocalization is quantified in ( M ). We tested colocalization between partners of the activated NOX2 complex in “M2” macrophages formed by membrane bound NOX2 and <t>p22</t> phox with p67 phox and p47 phox (p40 phox and rac1 are not labeled). It is known that p67 phox and p47 phox are located in the cytoplasm of the resting cells in agreement with the images shown in ( H ); ( I , J ) are p22 phox -p47 phox and p22 phox -p67 phox co-localization signals in macrophages without NS1, respectively; ( K , L ) are p22 phox -p47 phox and p22 phox -p67 phox co-localizations with NS1, respectively. Scale bars are 3 µm. Quantification of the 3D images is shown in ( M ) and was performed with Imaris 7.3 software ( http://www.bitplane.com/imaris , accessed on 6 February 2023).
    P22 Phox Ab75941, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p22 phox ab75941/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p22 phox ab75941 - by Bioz Stars, 2023-11
    86/100 stars

    Images

    1) Product Images from "Targeting M2 Macrophages with a Novel NADPH Oxidase Inhibitor"

    Article Title: Targeting M2 Macrophages with a Novel NADPH Oxidase Inhibitor

    Journal: Antioxidants

    doi: 10.3390/antiox12020440

    Confocal imaging of living cells (Left A – F) ( A , B ): monocytes isolated from blood of healthy donors; the red signal is the NOX2 and mitotracker in ( A , B ), respectively, the grey signal corresponds to the mitotracker and CellRox deep red reagent in ( A , B ), respectively; ( C , D ) Monocyte-derived macrophages (with CSF1-without NS1 treatment) using the same markers used for the living monocytes in ( A , B ); ( E , F ) the macrophages were treated with NS1 after their CSF1-induced differentiation from monocytes using the same markers as in ( A , B ); the green signal in ( F ) is the fluorescence of NS1; the ROS levels are quantified by the cell-permeable CellROX deep red reagent shown in light grey, the mitochondria are monitored by the red mitotracker reagent as quantified in ( G ); Co-localization experiments in fixed cells (Right H – L) The images ( H – L ) present co-localization experiments in fixed monocytes or CSF1-treated monocytes differentiated into macrophages . Each antibody is mentioned above each image, the merge, representing the colocalization is shown in yellow. The intensity of the colocalization is quantified in ( M ). We tested colocalization between partners of the activated NOX2 complex in “M2” macrophages formed by membrane bound NOX2 and p22 phox with p67 phox and p47 phox (p40 phox and rac1 are not labeled). It is known that p67 phox and p47 phox are located in the cytoplasm of the resting cells in agreement with the images shown in ( H ); ( I , J ) are p22 phox -p47 phox and p22 phox -p67 phox co-localization signals in macrophages without NS1, respectively; ( K , L ) are p22 phox -p47 phox and p22 phox -p67 phox co-localizations with NS1, respectively. Scale bars are 3 µm. Quantification of the 3D images is shown in ( M ) and was performed with Imaris 7.3 software ( http://www.bitplane.com/imaris , accessed on 6 February 2023).
    Figure Legend Snippet: Confocal imaging of living cells (Left A – F) ( A , B ): monocytes isolated from blood of healthy donors; the red signal is the NOX2 and mitotracker in ( A , B ), respectively, the grey signal corresponds to the mitotracker and CellRox deep red reagent in ( A , B ), respectively; ( C , D ) Monocyte-derived macrophages (with CSF1-without NS1 treatment) using the same markers used for the living monocytes in ( A , B ); ( E , F ) the macrophages were treated with NS1 after their CSF1-induced differentiation from monocytes using the same markers as in ( A , B ); the green signal in ( F ) is the fluorescence of NS1; the ROS levels are quantified by the cell-permeable CellROX deep red reagent shown in light grey, the mitochondria are monitored by the red mitotracker reagent as quantified in ( G ); Co-localization experiments in fixed cells (Right H – L) The images ( H – L ) present co-localization experiments in fixed monocytes or CSF1-treated monocytes differentiated into macrophages . Each antibody is mentioned above each image, the merge, representing the colocalization is shown in yellow. The intensity of the colocalization is quantified in ( M ). We tested colocalization between partners of the activated NOX2 complex in “M2” macrophages formed by membrane bound NOX2 and p22 phox with p67 phox and p47 phox (p40 phox and rac1 are not labeled). It is known that p67 phox and p47 phox are located in the cytoplasm of the resting cells in agreement with the images shown in ( H ); ( I , J ) are p22 phox -p47 phox and p22 phox -p67 phox co-localization signals in macrophages without NS1, respectively; ( K , L ) are p22 phox -p47 phox and p22 phox -p67 phox co-localizations with NS1, respectively. Scale bars are 3 µm. Quantification of the 3D images is shown in ( M ) and was performed with Imaris 7.3 software ( http://www.bitplane.com/imaris , accessed on 6 February 2023).

    Techniques Used: Imaging, Isolation, Derivative Assay, Fluorescence, Labeling, Software

    anti p22 phox antibody  (Thermo Fisher)


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    Thermo Fisher anti p22 phox antibody
    Optimized production of <t>NOX2/p22</t> <t>phox</t> liposomes using the cell-free expression system. Notes: ( A ) Effect of magnesium and potassium concentrations on cell-free production of <t>p22</t> phox protein. In vitro expression was performed in 96-well plates with a volume of 50 µL. ( B ) Effect of lipid composition on the expression of p22 phox subunit. Total protein fraction was obtained after the cell-free expression reaction, while the proteoliposome fraction was separated after a discontinuous sucrose gradient to separate them from liposomes and aggregated proteins. ( C ) Effect of the reaction time variation on NOX2 and p22 phox expression. Reactions were carried out at 30°C for 2 h, 4 h, 6 h or 16 h, in batch format (100 µL) and separated by SDS-PAGE on a 15% gel. P22 phox and NOX2 detection bands are indicated with stars. ( D ) Effect of the variation of iron and hemin concentration on protein expression. In vitro expression was performed in 96-well plates with a volume of 50 µL. In all experiments, monoclonal anti-His HRP-conjugated antibody and anti-NOX2 antibodies (clone 44.1) were used for the detection of p22 phox and NOX2, respectively. Optimal concentrations were indicated. * Indicates the location of NOX2 and p22 phox . Abbreviations: NTPs, nucleotide triphosphates; PLs, proteoliposomes; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.
    Anti P22 Phox Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p22 phox antibody/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p22 phox antibody - by Bioz Stars, 2023-11
    86/100 stars

    Images

    1) Product Images from "Therapeutic effects of proteoliposomes on X-linked chronic granulomatous disease: proof of concept using macrophages differentiated from patient-specific induced pluripotent stem cells"

    Article Title: Therapeutic effects of proteoliposomes on X-linked chronic granulomatous disease: proof of concept using macrophages differentiated from patient-specific induced pluripotent stem cells

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S128611

    Optimized production of NOX2/p22 phox liposomes using the cell-free expression system. Notes: ( A ) Effect of magnesium and potassium concentrations on cell-free production of p22 phox protein. In vitro expression was performed in 96-well plates with a volume of 50 µL. ( B ) Effect of lipid composition on the expression of p22 phox subunit. Total protein fraction was obtained after the cell-free expression reaction, while the proteoliposome fraction was separated after a discontinuous sucrose gradient to separate them from liposomes and aggregated proteins. ( C ) Effect of the reaction time variation on NOX2 and p22 phox expression. Reactions were carried out at 30°C for 2 h, 4 h, 6 h or 16 h, in batch format (100 µL) and separated by SDS-PAGE on a 15% gel. P22 phox and NOX2 detection bands are indicated with stars. ( D ) Effect of the variation of iron and hemin concentration on protein expression. In vitro expression was performed in 96-well plates with a volume of 50 µL. In all experiments, monoclonal anti-His HRP-conjugated antibody and anti-NOX2 antibodies (clone 44.1) were used for the detection of p22 phox and NOX2, respectively. Optimal concentrations were indicated. * Indicates the location of NOX2 and p22 phox . Abbreviations: NTPs, nucleotide triphosphates; PLs, proteoliposomes; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.
    Figure Legend Snippet: Optimized production of NOX2/p22 phox liposomes using the cell-free expression system. Notes: ( A ) Effect of magnesium and potassium concentrations on cell-free production of p22 phox protein. In vitro expression was performed in 96-well plates with a volume of 50 µL. ( B ) Effect of lipid composition on the expression of p22 phox subunit. Total protein fraction was obtained after the cell-free expression reaction, while the proteoliposome fraction was separated after a discontinuous sucrose gradient to separate them from liposomes and aggregated proteins. ( C ) Effect of the reaction time variation on NOX2 and p22 phox expression. Reactions were carried out at 30°C for 2 h, 4 h, 6 h or 16 h, in batch format (100 µL) and separated by SDS-PAGE on a 15% gel. P22 phox and NOX2 detection bands are indicated with stars. ( D ) Effect of the variation of iron and hemin concentration on protein expression. In vitro expression was performed in 96-well plates with a volume of 50 µL. In all experiments, monoclonal anti-His HRP-conjugated antibody and anti-NOX2 antibodies (clone 44.1) were used for the detection of p22 phox and NOX2, respectively. Optimal concentrations were indicated. * Indicates the location of NOX2 and p22 phox . Abbreviations: NTPs, nucleotide triphosphates; PLs, proteoliposomes; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.

    Techniques Used: Expressing, In Vitro, SDS Page, Concentration Assay, Polyacrylamide Gel Electrophoresis

    In vitro physicochemical characterization of NOX2/p22 phox and negative liposomes. Notes: ( A ) Particle size distribution of NOX2/p22 phox and negative liposomes by DLS. ( B ) Coomassie blue staining of NOX2/p22 phox and negative liposomes (The * and ▲ indicate the location of NOX2 and p22 phox respectively) and Western blot analysis of NOX2/p22 phox and negative liposomes using monoclonal antibodies against NOX2 and p22 phox . ( C ) Dithionite-reduced minus-oxidized spectra of NOX2/p22 phox and negative liposomes. Abbreviations: DLS, dynamic light scattering; OD, optical density; PLs, proteoliposomes.
    Figure Legend Snippet: In vitro physicochemical characterization of NOX2/p22 phox and negative liposomes. Notes: ( A ) Particle size distribution of NOX2/p22 phox and negative liposomes by DLS. ( B ) Coomassie blue staining of NOX2/p22 phox and negative liposomes (The * and ▲ indicate the location of NOX2 and p22 phox respectively) and Western blot analysis of NOX2/p22 phox and negative liposomes using monoclonal antibodies against NOX2 and p22 phox . ( C ) Dithionite-reduced minus-oxidized spectra of NOX2/p22 phox and negative liposomes. Abbreviations: DLS, dynamic light scattering; OD, optical density; PLs, proteoliposomes.

    Techniques Used: In Vitro, Staining, Western Blot

    In vitro NADPH oxidase activity of NOX2/p22 phox and negative liposomes. Notes: ( A ) Representative results of the NADPH oxidase activity of NOX2/p22 phox (8 pmol cytochrome b 558 ) and negative liposomes in a cell-free system assay in the presence of the recombinant p47 phox , p67 phox and Rac (1 µM), arachidonic acid (400 µM) and NBT (100 µM), stimulated or not with NADPH (200 µM). ( B ) NADPH oxidase activity was expressed in moles of superoxide O 2 •− produced/s/mol of heme and measured before (total activity) or after SOD addition (SOD non-inhibitable activity). Cell-free system assay was performed in the same experimental conditions as ( A ). Abbreviations: DPI, diphenyleneiodonium; NADPH, nicotinamide adenine dinucleotide phosphate; NBT, nitroblue tetrazolium; OD, optical density; O 2 •− , superoxide anion; SOD, superoxide dismutase; PLs, proteoliposomes.
    Figure Legend Snippet: In vitro NADPH oxidase activity of NOX2/p22 phox and negative liposomes. Notes: ( A ) Representative results of the NADPH oxidase activity of NOX2/p22 phox (8 pmol cytochrome b 558 ) and negative liposomes in a cell-free system assay in the presence of the recombinant p47 phox , p67 phox and Rac (1 µM), arachidonic acid (400 µM) and NBT (100 µM), stimulated or not with NADPH (200 µM). ( B ) NADPH oxidase activity was expressed in moles of superoxide O 2 •− produced/s/mol of heme and measured before (total activity) or after SOD addition (SOD non-inhibitable activity). Cell-free system assay was performed in the same experimental conditions as ( A ). Abbreviations: DPI, diphenyleneiodonium; NADPH, nicotinamide adenine dinucleotide phosphate; NBT, nitroblue tetrazolium; OD, optical density; O 2 •− , superoxide anion; SOD, superoxide dismutase; PLs, proteoliposomes.

    Techniques Used: In Vitro, Activity Assay, Recombinant, Produced

    Analysis of the membrane delivery of NOX2 and p22 phox subunits in X 0 -CGD iPSC-derived macrophages after NOX2/p22 phox liposome treatment. Notes: ( A ) Flow cytometry analysis of NOX2 and p22 phox expression using monoclonal antibodies in WT and untreated X 0 -CGD macrophages (black curve), and X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox (red curve) or negative (green curve) liposomes. Isotype controls are represented by gray-filled curves. MFIs were indicated for each condition, and the shift of fluorescence was calculated as the percentage of increased fluorescence compared to untreated macrophages. ( B ) Confocal microscopy images showing the staining of NOX2 subunit with 7D5 antibody and AF488-conjugated secondary antibody (green) in WT and X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox or negative liposomes. Nuclei were counterstained with Hoechst 33258 (blue); scale bars =20 µm. The same observations were obtained in at least two experiments. Abbreviations: CGD, chronic granulomatous disease; iPSC, induced pluripotent stem cell; MFIs, mean fluorescence intensities; MΦ, macrophages; WT, wild type; X 0 -CGD, X 0 -linked CGD; XCGD, X-linked CGD.
    Figure Legend Snippet: Analysis of the membrane delivery of NOX2 and p22 phox subunits in X 0 -CGD iPSC-derived macrophages after NOX2/p22 phox liposome treatment. Notes: ( A ) Flow cytometry analysis of NOX2 and p22 phox expression using monoclonal antibodies in WT and untreated X 0 -CGD macrophages (black curve), and X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox (red curve) or negative (green curve) liposomes. Isotype controls are represented by gray-filled curves. MFIs were indicated for each condition, and the shift of fluorescence was calculated as the percentage of increased fluorescence compared to untreated macrophages. ( B ) Confocal microscopy images showing the staining of NOX2 subunit with 7D5 antibody and AF488-conjugated secondary antibody (green) in WT and X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox or negative liposomes. Nuclei were counterstained with Hoechst 33258 (blue); scale bars =20 µm. The same observations were obtained in at least two experiments. Abbreviations: CGD, chronic granulomatous disease; iPSC, induced pluripotent stem cell; MFIs, mean fluorescence intensities; MΦ, macrophages; WT, wild type; X 0 -CGD, X 0 -linked CGD; XCGD, X-linked CGD.

    Techniques Used: Derivative Assay, Flow Cytometry, Expressing, Fluorescence, Confocal Microscopy, Staining

    Location of NOX2 in liposome-treated X 0 -CGD iPSC-derived macrophages after C . albicans phagocytosis. Notes: ( A ) Confocal microscopy images showing the staining of NOX2 subunit with 7D5 antibody and AF488-conjugated secondary antibody (green) in X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox or negative liposomes and then for 4 h with C . albicans strain at an MOI of 3:1. ( B ) Z-stack images (top to bottom of cell) of an NOX2/p22 phox liposome-treated X 0 -CGD macrophage incubated for 4 h with C . albicans strain at an MOI of 3:1. Nuclei were counterstained with Hoechst 33258 (blue); scale bars =20 µm in ( A ) and 10 µm in ( B ). The same observations were obtained in at least two experiments. Abbreviations: C. albicans; Candida albicans , CGD, chronic granulomatous disease; iPSC, induced pluripotent stem cell; MOI, multiplicity of infection; X 0 -CGD, X 0 -linked CGD; XCGD, X-linked CGD.
    Figure Legend Snippet: Location of NOX2 in liposome-treated X 0 -CGD iPSC-derived macrophages after C . albicans phagocytosis. Notes: ( A ) Confocal microscopy images showing the staining of NOX2 subunit with 7D5 antibody and AF488-conjugated secondary antibody (green) in X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox or negative liposomes and then for 4 h with C . albicans strain at an MOI of 3:1. ( B ) Z-stack images (top to bottom of cell) of an NOX2/p22 phox liposome-treated X 0 -CGD macrophage incubated for 4 h with C . albicans strain at an MOI of 3:1. Nuclei were counterstained with Hoechst 33258 (blue); scale bars =20 µm in ( A ) and 10 µm in ( B ). The same observations were obtained in at least two experiments. Abbreviations: C. albicans; Candida albicans , CGD, chronic granulomatous disease; iPSC, induced pluripotent stem cell; MOI, multiplicity of infection; X 0 -CGD, X 0 -linked CGD; XCGD, X-linked CGD.

    Techniques Used: Derivative Assay, Confocal Microscopy, Staining, Incubation, Infection

    Analysis of in cellulo toxicity and NADPH oxidase activity restoration in X 0 -CGD iPSC-derived macrophages. Notes: ( A ) Viability of CGDX 0 iPSC-derived macrophages after 8, 12 and 24 h of treatment. Untreated cells are used as positive control (100%) and DMSO-treated cells as negative control. Data are expressed as mean ± SEM ( n =3). The differences of viability between the untreated cells and the liposome-treated cells were analyzed using the nonparametric Mann–Whitney test. ( B ) Morphological images of X 0 -CGD iPSC-derived macrophages after 8, 12 and 24 h of treatment with NOX2/p22 phox liposomes or incubated with IMDM only (magnification ×10). ( C ) X 0 -CGD macrophages were treated for 8 h with NOX2/p22 phox or negative liposomes. Then, WT and X 0 -CGD macrophages were stimulated with PMA (resting; scale bars =50 µm). Abbreviations: CGD, chronic granulomatous disease; DMSO, dimethylsulfoxide; IMDM, Iscove’s Modified Dulbecco’s Medium; iPSC, induced pluripotent stem cell; NADPH, nicotinamide adenine dinucleotide phosphate; ns, nonsignificant; PMA, phorbolmyristate-acetate; SEM, standard error of the mean; WT, wild type; X 0 -CGD, X 0 -linked CGD.
    Figure Legend Snippet: Analysis of in cellulo toxicity and NADPH oxidase activity restoration in X 0 -CGD iPSC-derived macrophages. Notes: ( A ) Viability of CGDX 0 iPSC-derived macrophages after 8, 12 and 24 h of treatment. Untreated cells are used as positive control (100%) and DMSO-treated cells as negative control. Data are expressed as mean ± SEM ( n =3). The differences of viability between the untreated cells and the liposome-treated cells were analyzed using the nonparametric Mann–Whitney test. ( B ) Morphological images of X 0 -CGD iPSC-derived macrophages after 8, 12 and 24 h of treatment with NOX2/p22 phox liposomes or incubated with IMDM only (magnification ×10). ( C ) X 0 -CGD macrophages were treated for 8 h with NOX2/p22 phox or negative liposomes. Then, WT and X 0 -CGD macrophages were stimulated with PMA (resting; scale bars =50 µm). Abbreviations: CGD, chronic granulomatous disease; DMSO, dimethylsulfoxide; IMDM, Iscove’s Modified Dulbecco’s Medium; iPSC, induced pluripotent stem cell; NADPH, nicotinamide adenine dinucleotide phosphate; ns, nonsignificant; PMA, phorbolmyristate-acetate; SEM, standard error of the mean; WT, wild type; X 0 -CGD, X 0 -linked CGD.

    Techniques Used: Activity Assay, Derivative Assay, Positive Control, Negative Control, MANN-WHITNEY, Incubation, Modification

    anti p22 phox  (Thermo Fisher)


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    Thermo Fisher anti p22 phox
    Anti P22 Phox, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p22 phox/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p22 phox - by Bioz Stars, 2023-11
    86/100 stars

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    p22 phox  (Thermo Fisher)


    Bioz Verified Symbol Thermo Fisher is a verified supplier
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    Thermo Fisher p22 phox
    P22 Phox, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p22 phox/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p22 phox - by Bioz Stars, 2023-11
    86/100 stars

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    p22 phox  (Thermo Fisher)


    Bioz Verified Symbol Thermo Fisher is a verified supplier
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    Thermo Fisher p22 phox
    P22 Phox, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p22 phox/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p22 phox - by Bioz Stars, 2023-11
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    p22 phox  (Thermo Fisher)


    Bioz Verified Symbol Thermo Fisher is a verified supplier
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    Thermo Fisher p22 phox
    Effect of ursolic acid (UA) on the NOX subunits and Hh signaling pathway during quiescent hepatic stellate cell (HSC) activation. Quiescent HSCs were stimulated with transforming growth factor (TGF)-β1 (5 µg/l) for 24 h. Protein expression levels of gp91 <t>phox</t> , p67 phox , <t>p22</t> phox , Rac1, Shh, Smo, and Gli2 were significantly higher than in the control group. Treatment with UA (40 µM) inhibited expression of these proteins. UA and DPI had comparable effects. DPI, diphenyleneiodonium chloride; Shh, sonic hedgehog; Smo, sterol-4-alpha-methyl oxidase; Gli2, Gli family zinc finger 2.
    P22 Phox, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p22 phox/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p22 phox - by Bioz Stars, 2023-11
    86/100 stars

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    1) Product Images from "Ursolic acid suppresses TGF-β1-induced quiescent HSC activation and transformation by inhibiting NADPH oxidase expression and Hedgehog signaling"

    Article Title: Ursolic acid suppresses TGF-β1-induced quiescent HSC activation and transformation by inhibiting NADPH oxidase expression and Hedgehog signaling

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2017.5001

    Effect of ursolic acid (UA) on the NOX subunits and Hh signaling pathway during quiescent hepatic stellate cell (HSC) activation. Quiescent HSCs were stimulated with transforming growth factor (TGF)-β1 (5 µg/l) for 24 h. Protein expression levels of gp91 phox , p67 phox , p22 phox , Rac1, Shh, Smo, and Gli2 were significantly higher than in the control group. Treatment with UA (40 µM) inhibited expression of these proteins. UA and DPI had comparable effects. DPI, diphenyleneiodonium chloride; Shh, sonic hedgehog; Smo, sterol-4-alpha-methyl oxidase; Gli2, Gli family zinc finger 2.
    Figure Legend Snippet: Effect of ursolic acid (UA) on the NOX subunits and Hh signaling pathway during quiescent hepatic stellate cell (HSC) activation. Quiescent HSCs were stimulated with transforming growth factor (TGF)-β1 (5 µg/l) for 24 h. Protein expression levels of gp91 phox , p67 phox , p22 phox , Rac1, Shh, Smo, and Gli2 were significantly higher than in the control group. Treatment with UA (40 µM) inhibited expression of these proteins. UA and DPI had comparable effects. DPI, diphenyleneiodonium chloride; Shh, sonic hedgehog; Smo, sterol-4-alpha-methyl oxidase; Gli2, Gli family zinc finger 2.

    Techniques Used: Activation Assay, Expressing

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    • p22 phox ab75941  (Thermo Fisher)
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    Thermo Fisher p22 phox ab75941
    Confocal imaging of living cells (Left A – F) ( A , B ): monocytes isolated from blood of healthy donors; the red signal is the NOX2 and mitotracker in ( A , B ), respectively, the grey signal corresponds to the mitotracker and CellRox deep red reagent in ( A , B ), respectively; ( C , D ) Monocyte-derived macrophages (with CSF1-without NS1 treatment) using the same markers used for the living monocytes in ( A , B ); ( E , F ) the macrophages were treated with NS1 after their CSF1-induced differentiation from monocytes using the same markers as in ( A , B ); the green signal in ( F ) is the fluorescence of NS1; the ROS levels are quantified by the cell-permeable CellROX deep red reagent shown in light grey, the mitochondria are monitored by the red mitotracker reagent as quantified in ( G ); Co-localization experiments in fixed cells (Right H – L) The images ( H – L ) present co-localization experiments in fixed monocytes or CSF1-treated monocytes differentiated into macrophages . Each antibody is mentioned above each image, the merge, representing the colocalization is shown in yellow. The intensity of the colocalization is quantified in ( M ). We tested colocalization between partners of the activated NOX2 complex in “M2” macrophages formed by membrane bound NOX2 and <t>p22</t> phox with p67 phox and p47 phox (p40 phox and rac1 are not labeled). It is known that p67 phox and p47 phox are located in the cytoplasm of the resting cells in agreement with the images shown in ( H ); ( I , J ) are p22 phox -p47 phox and p22 phox -p67 phox co-localization signals in macrophages without NS1, respectively; ( K , L ) are p22 phox -p47 phox and p22 phox -p67 phox co-localizations with NS1, respectively. Scale bars are 3 µm. Quantification of the 3D images is shown in ( M ) and was performed with Imaris 7.3 software ( http://www.bitplane.com/imaris , accessed on 6 February 2023).
    P22 Phox Ab75941, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p22 phox ab75941/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p22 phox ab75941 - by Bioz Stars, 2023-11
    86/100 stars
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    anti p22 phox antibody  (Thermo Fisher)
    86
    Thermo Fisher anti p22 phox antibody
    Optimized production of <t>NOX2/p22</t> <t>phox</t> liposomes using the cell-free expression system. Notes: ( A ) Effect of magnesium and potassium concentrations on cell-free production of <t>p22</t> phox protein. In vitro expression was performed in 96-well plates with a volume of 50 µL. ( B ) Effect of lipid composition on the expression of p22 phox subunit. Total protein fraction was obtained after the cell-free expression reaction, while the proteoliposome fraction was separated after a discontinuous sucrose gradient to separate them from liposomes and aggregated proteins. ( C ) Effect of the reaction time variation on NOX2 and p22 phox expression. Reactions were carried out at 30°C for 2 h, 4 h, 6 h or 16 h, in batch format (100 µL) and separated by SDS-PAGE on a 15% gel. P22 phox and NOX2 detection bands are indicated with stars. ( D ) Effect of the variation of iron and hemin concentration on protein expression. In vitro expression was performed in 96-well plates with a volume of 50 µL. In all experiments, monoclonal anti-His HRP-conjugated antibody and anti-NOX2 antibodies (clone 44.1) were used for the detection of p22 phox and NOX2, respectively. Optimal concentrations were indicated. * Indicates the location of NOX2 and p22 phox . Abbreviations: NTPs, nucleotide triphosphates; PLs, proteoliposomes; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.
    Anti P22 Phox Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p22 phox antibody/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p22 phox antibody - by Bioz Stars, 2023-11
    86/100 stars
    		  Buy from Supplier
    anti p22 phox  (Thermo Fisher)
    86
    Thermo Fisher anti p22 phox
    Optimized production of <t>NOX2/p22</t> <t>phox</t> liposomes using the cell-free expression system. Notes: ( A ) Effect of magnesium and potassium concentrations on cell-free production of <t>p22</t> phox protein. In vitro expression was performed in 96-well plates with a volume of 50 µL. ( B ) Effect of lipid composition on the expression of p22 phox subunit. Total protein fraction was obtained after the cell-free expression reaction, while the proteoliposome fraction was separated after a discontinuous sucrose gradient to separate them from liposomes and aggregated proteins. ( C ) Effect of the reaction time variation on NOX2 and p22 phox expression. Reactions were carried out at 30°C for 2 h, 4 h, 6 h or 16 h, in batch format (100 µL) and separated by SDS-PAGE on a 15% gel. P22 phox and NOX2 detection bands are indicated with stars. ( D ) Effect of the variation of iron and hemin concentration on protein expression. In vitro expression was performed in 96-well plates with a volume of 50 µL. In all experiments, monoclonal anti-His HRP-conjugated antibody and anti-NOX2 antibodies (clone 44.1) were used for the detection of p22 phox and NOX2, respectively. Optimal concentrations were indicated. * Indicates the location of NOX2 and p22 phox . Abbreviations: NTPs, nucleotide triphosphates; PLs, proteoliposomes; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.
    Anti P22 Phox, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p22 phox/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p22 phox - by Bioz Stars, 2023-11
    86/100 stars
    		  Buy from Supplier
    p22 phox  (Thermo Fisher)
    86
    Thermo Fisher p22 phox
    Optimized production of <t>NOX2/p22</t> <t>phox</t> liposomes using the cell-free expression system. Notes: ( A ) Effect of magnesium and potassium concentrations on cell-free production of <t>p22</t> phox protein. In vitro expression was performed in 96-well plates with a volume of 50 µL. ( B ) Effect of lipid composition on the expression of p22 phox subunit. Total protein fraction was obtained after the cell-free expression reaction, while the proteoliposome fraction was separated after a discontinuous sucrose gradient to separate them from liposomes and aggregated proteins. ( C ) Effect of the reaction time variation on NOX2 and p22 phox expression. Reactions were carried out at 30°C for 2 h, 4 h, 6 h or 16 h, in batch format (100 µL) and separated by SDS-PAGE on a 15% gel. P22 phox and NOX2 detection bands are indicated with stars. ( D ) Effect of the variation of iron and hemin concentration on protein expression. In vitro expression was performed in 96-well plates with a volume of 50 µL. In all experiments, monoclonal anti-His HRP-conjugated antibody and anti-NOX2 antibodies (clone 44.1) were used for the detection of p22 phox and NOX2, respectively. Optimal concentrations were indicated. * Indicates the location of NOX2 and p22 phox . Abbreviations: NTPs, nucleotide triphosphates; PLs, proteoliposomes; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.
    P22 Phox, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p22 phox/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p22 phox - by Bioz Stars, 2023-11
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    Image Search Results


    Confocal imaging of living cells (Left A – F) ( A , B ): monocytes isolated from blood of healthy donors; the red signal is the NOX2 and mitotracker in ( A , B ), respectively, the grey signal corresponds to the mitotracker and CellRox deep red reagent in ( A , B ), respectively; ( C , D ) Monocyte-derived macrophages (with CSF1-without NS1 treatment) using the same markers used for the living monocytes in ( A , B ); ( E , F ) the macrophages were treated with NS1 after their CSF1-induced differentiation from monocytes using the same markers as in ( A , B ); the green signal in ( F ) is the fluorescence of NS1; the ROS levels are quantified by the cell-permeable CellROX deep red reagent shown in light grey, the mitochondria are monitored by the red mitotracker reagent as quantified in ( G ); Co-localization experiments in fixed cells (Right H – L) The images ( H – L ) present co-localization experiments in fixed monocytes or CSF1-treated monocytes differentiated into macrophages . Each antibody is mentioned above each image, the merge, representing the colocalization is shown in yellow. The intensity of the colocalization is quantified in ( M ). We tested colocalization between partners of the activated NOX2 complex in “M2” macrophages formed by membrane bound NOX2 and p22 phox with p67 phox and p47 phox (p40 phox and rac1 are not labeled). It is known that p67 phox and p47 phox are located in the cytoplasm of the resting cells in agreement with the images shown in ( H ); ( I , J ) are p22 phox -p47 phox and p22 phox -p67 phox co-localization signals in macrophages without NS1, respectively; ( K , L ) are p22 phox -p47 phox and p22 phox -p67 phox co-localizations with NS1, respectively. Scale bars are 3 µm. Quantification of the 3D images is shown in ( M ) and was performed with Imaris 7.3 software ( http://www.bitplane.com/imaris , accessed on 6 February 2023).

    Journal: Antioxidants

    Article Title: Targeting M2 Macrophages with a Novel NADPH Oxidase Inhibitor

    doi: 10.3390/antiox12020440

    Figure Lengend Snippet: Confocal imaging of living cells (Left A – F) ( A , B ): monocytes isolated from blood of healthy donors; the red signal is the NOX2 and mitotracker in ( A , B ), respectively, the grey signal corresponds to the mitotracker and CellRox deep red reagent in ( A , B ), respectively; ( C , D ) Monocyte-derived macrophages (with CSF1-without NS1 treatment) using the same markers used for the living monocytes in ( A , B ); ( E , F ) the macrophages were treated with NS1 after their CSF1-induced differentiation from monocytes using the same markers as in ( A , B ); the green signal in ( F ) is the fluorescence of NS1; the ROS levels are quantified by the cell-permeable CellROX deep red reagent shown in light grey, the mitochondria are monitored by the red mitotracker reagent as quantified in ( G ); Co-localization experiments in fixed cells (Right H – L) The images ( H – L ) present co-localization experiments in fixed monocytes or CSF1-treated monocytes differentiated into macrophages . Each antibody is mentioned above each image, the merge, representing the colocalization is shown in yellow. The intensity of the colocalization is quantified in ( M ). We tested colocalization between partners of the activated NOX2 complex in “M2” macrophages formed by membrane bound NOX2 and p22 phox with p67 phox and p47 phox (p40 phox and rac1 are not labeled). It is known that p67 phox and p47 phox are located in the cytoplasm of the resting cells in agreement with the images shown in ( H ); ( I , J ) are p22 phox -p47 phox and p22 phox -p67 phox co-localization signals in macrophages without NS1, respectively; ( K , L ) are p22 phox -p47 phox and p22 phox -p67 phox co-localizations with NS1, respectively. Scale bars are 3 µm. Quantification of the 3D images is shown in ( M ) and was performed with Imaris 7.3 software ( http://www.bitplane.com/imaris , accessed on 6 February 2023).

    Article Snippet: Mitotracker (red or far-red) and CellRox deep red were purchased from Thermo-Fischer and Invitrogen (Les Ulys, France), respectively; antibodies against NOX2 (ab80897) and p22 phox (ab75941) were acquired from Abcam; p47 phox (sc7660) and p67 phox (sc7663) were acquired from Santa Cruz; and NS1 was synthesized by Innoverda according to the published synthesis [ ].

    Techniques: Imaging, Isolation, Derivative Assay, Fluorescence, Labeling, Software

    Optimized production of NOX2/p22 phox liposomes using the cell-free expression system. Notes: ( A ) Effect of magnesium and potassium concentrations on cell-free production of p22 phox protein. In vitro expression was performed in 96-well plates with a volume of 50 µL. ( B ) Effect of lipid composition on the expression of p22 phox subunit. Total protein fraction was obtained after the cell-free expression reaction, while the proteoliposome fraction was separated after a discontinuous sucrose gradient to separate them from liposomes and aggregated proteins. ( C ) Effect of the reaction time variation on NOX2 and p22 phox expression. Reactions were carried out at 30°C for 2 h, 4 h, 6 h or 16 h, in batch format (100 µL) and separated by SDS-PAGE on a 15% gel. P22 phox and NOX2 detection bands are indicated with stars. ( D ) Effect of the variation of iron and hemin concentration on protein expression. In vitro expression was performed in 96-well plates with a volume of 50 µL. In all experiments, monoclonal anti-His HRP-conjugated antibody and anti-NOX2 antibodies (clone 44.1) were used for the detection of p22 phox and NOX2, respectively. Optimal concentrations were indicated. * Indicates the location of NOX2 and p22 phox . Abbreviations: NTPs, nucleotide triphosphates; PLs, proteoliposomes; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.

    Journal: International Journal of Nanomedicine

    Article Title: Therapeutic effects of proteoliposomes on X-linked chronic granulomatous disease: proof of concept using macrophages differentiated from patient-specific induced pluripotent stem cells

    doi: 10.2147/IJN.S128611

    Figure Lengend Snippet: Optimized production of NOX2/p22 phox liposomes using the cell-free expression system. Notes: ( A ) Effect of magnesium and potassium concentrations on cell-free production of p22 phox protein. In vitro expression was performed in 96-well plates with a volume of 50 µL. ( B ) Effect of lipid composition on the expression of p22 phox subunit. Total protein fraction was obtained after the cell-free expression reaction, while the proteoliposome fraction was separated after a discontinuous sucrose gradient to separate them from liposomes and aggregated proteins. ( C ) Effect of the reaction time variation on NOX2 and p22 phox expression. Reactions were carried out at 30°C for 2 h, 4 h, 6 h or 16 h, in batch format (100 µL) and separated by SDS-PAGE on a 15% gel. P22 phox and NOX2 detection bands are indicated with stars. ( D ) Effect of the variation of iron and hemin concentration on protein expression. In vitro expression was performed in 96-well plates with a volume of 50 µL. In all experiments, monoclonal anti-His HRP-conjugated antibody and anti-NOX2 antibodies (clone 44.1) were used for the detection of p22 phox and NOX2, respectively. Optimal concentrations were indicated. * Indicates the location of NOX2 and p22 phox . Abbreviations: NTPs, nucleotide triphosphates; PLs, proteoliposomes; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.

    Article Snippet: Paraformaldehyde (PFA), fetal bovine serum (FBS), goat serum, Iscove’s Modified Dulbecco’s Medium (IMDM), Alexa Fluor ® 488-conjugated goat-F(ab′) 2 fragment anti-mouse IgG1 (H+L), anti-NOX2 antibody (clone 54.1) and anti-p22 phox antibody (clone 44.1) were from Thermo Fisher Scientific (Courtaboeuf, France).

    Techniques: Expressing, In Vitro, SDS Page, Concentration Assay, Polyacrylamide Gel Electrophoresis

    In vitro physicochemical characterization of NOX2/p22 phox and negative liposomes. Notes: ( A ) Particle size distribution of NOX2/p22 phox and negative liposomes by DLS. ( B ) Coomassie blue staining of NOX2/p22 phox and negative liposomes (The * and ▲ indicate the location of NOX2 and p22 phox respectively) and Western blot analysis of NOX2/p22 phox and negative liposomes using monoclonal antibodies against NOX2 and p22 phox . ( C ) Dithionite-reduced minus-oxidized spectra of NOX2/p22 phox and negative liposomes. Abbreviations: DLS, dynamic light scattering; OD, optical density; PLs, proteoliposomes.

    Journal: International Journal of Nanomedicine

    Article Title: Therapeutic effects of proteoliposomes on X-linked chronic granulomatous disease: proof of concept using macrophages differentiated from patient-specific induced pluripotent stem cells

    doi: 10.2147/IJN.S128611

    Figure Lengend Snippet: In vitro physicochemical characterization of NOX2/p22 phox and negative liposomes. Notes: ( A ) Particle size distribution of NOX2/p22 phox and negative liposomes by DLS. ( B ) Coomassie blue staining of NOX2/p22 phox and negative liposomes (The * and ▲ indicate the location of NOX2 and p22 phox respectively) and Western blot analysis of NOX2/p22 phox and negative liposomes using monoclonal antibodies against NOX2 and p22 phox . ( C ) Dithionite-reduced minus-oxidized spectra of NOX2/p22 phox and negative liposomes. Abbreviations: DLS, dynamic light scattering; OD, optical density; PLs, proteoliposomes.

    Article Snippet: Paraformaldehyde (PFA), fetal bovine serum (FBS), goat serum, Iscove’s Modified Dulbecco’s Medium (IMDM), Alexa Fluor ® 488-conjugated goat-F(ab′) 2 fragment anti-mouse IgG1 (H+L), anti-NOX2 antibody (clone 54.1) and anti-p22 phox antibody (clone 44.1) were from Thermo Fisher Scientific (Courtaboeuf, France).

    Techniques: In Vitro, Staining, Western Blot

    In vitro NADPH oxidase activity of NOX2/p22 phox and negative liposomes. Notes: ( A ) Representative results of the NADPH oxidase activity of NOX2/p22 phox (8 pmol cytochrome b 558 ) and negative liposomes in a cell-free system assay in the presence of the recombinant p47 phox , p67 phox and Rac (1 µM), arachidonic acid (400 µM) and NBT (100 µM), stimulated or not with NADPH (200 µM). ( B ) NADPH oxidase activity was expressed in moles of superoxide O 2 •− produced/s/mol of heme and measured before (total activity) or after SOD addition (SOD non-inhibitable activity). Cell-free system assay was performed in the same experimental conditions as ( A ). Abbreviations: DPI, diphenyleneiodonium; NADPH, nicotinamide adenine dinucleotide phosphate; NBT, nitroblue tetrazolium; OD, optical density; O 2 •− , superoxide anion; SOD, superoxide dismutase; PLs, proteoliposomes.

    Journal: International Journal of Nanomedicine

    Article Title: Therapeutic effects of proteoliposomes on X-linked chronic granulomatous disease: proof of concept using macrophages differentiated from patient-specific induced pluripotent stem cells

    doi: 10.2147/IJN.S128611

    Figure Lengend Snippet: In vitro NADPH oxidase activity of NOX2/p22 phox and negative liposomes. Notes: ( A ) Representative results of the NADPH oxidase activity of NOX2/p22 phox (8 pmol cytochrome b 558 ) and negative liposomes in a cell-free system assay in the presence of the recombinant p47 phox , p67 phox and Rac (1 µM), arachidonic acid (400 µM) and NBT (100 µM), stimulated or not with NADPH (200 µM). ( B ) NADPH oxidase activity was expressed in moles of superoxide O 2 •− produced/s/mol of heme and measured before (total activity) or after SOD addition (SOD non-inhibitable activity). Cell-free system assay was performed in the same experimental conditions as ( A ). Abbreviations: DPI, diphenyleneiodonium; NADPH, nicotinamide adenine dinucleotide phosphate; NBT, nitroblue tetrazolium; OD, optical density; O 2 •− , superoxide anion; SOD, superoxide dismutase; PLs, proteoliposomes.

    Article Snippet: Paraformaldehyde (PFA), fetal bovine serum (FBS), goat serum, Iscove’s Modified Dulbecco’s Medium (IMDM), Alexa Fluor ® 488-conjugated goat-F(ab′) 2 fragment anti-mouse IgG1 (H+L), anti-NOX2 antibody (clone 54.1) and anti-p22 phox antibody (clone 44.1) were from Thermo Fisher Scientific (Courtaboeuf, France).

    Techniques: In Vitro, Activity Assay, Recombinant, Produced

    Analysis of the membrane delivery of NOX2 and p22 phox subunits in X 0 -CGD iPSC-derived macrophages after NOX2/p22 phox liposome treatment. Notes: ( A ) Flow cytometry analysis of NOX2 and p22 phox expression using monoclonal antibodies in WT and untreated X 0 -CGD macrophages (black curve), and X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox (red curve) or negative (green curve) liposomes. Isotype controls are represented by gray-filled curves. MFIs were indicated for each condition, and the shift of fluorescence was calculated as the percentage of increased fluorescence compared to untreated macrophages. ( B ) Confocal microscopy images showing the staining of NOX2 subunit with 7D5 antibody and AF488-conjugated secondary antibody (green) in WT and X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox or negative liposomes. Nuclei were counterstained with Hoechst 33258 (blue); scale bars =20 µm. The same observations were obtained in at least two experiments. Abbreviations: CGD, chronic granulomatous disease; iPSC, induced pluripotent stem cell; MFIs, mean fluorescence intensities; MΦ, macrophages; WT, wild type; X 0 -CGD, X 0 -linked CGD; XCGD, X-linked CGD.

    Journal: International Journal of Nanomedicine

    Article Title: Therapeutic effects of proteoliposomes on X-linked chronic granulomatous disease: proof of concept using macrophages differentiated from patient-specific induced pluripotent stem cells

    doi: 10.2147/IJN.S128611

    Figure Lengend Snippet: Analysis of the membrane delivery of NOX2 and p22 phox subunits in X 0 -CGD iPSC-derived macrophages after NOX2/p22 phox liposome treatment. Notes: ( A ) Flow cytometry analysis of NOX2 and p22 phox expression using monoclonal antibodies in WT and untreated X 0 -CGD macrophages (black curve), and X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox (red curve) or negative (green curve) liposomes. Isotype controls are represented by gray-filled curves. MFIs were indicated for each condition, and the shift of fluorescence was calculated as the percentage of increased fluorescence compared to untreated macrophages. ( B ) Confocal microscopy images showing the staining of NOX2 subunit with 7D5 antibody and AF488-conjugated secondary antibody (green) in WT and X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox or negative liposomes. Nuclei were counterstained with Hoechst 33258 (blue); scale bars =20 µm. The same observations were obtained in at least two experiments. Abbreviations: CGD, chronic granulomatous disease; iPSC, induced pluripotent stem cell; MFIs, mean fluorescence intensities; MΦ, macrophages; WT, wild type; X 0 -CGD, X 0 -linked CGD; XCGD, X-linked CGD.

    Article Snippet: Paraformaldehyde (PFA), fetal bovine serum (FBS), goat serum, Iscove’s Modified Dulbecco’s Medium (IMDM), Alexa Fluor ® 488-conjugated goat-F(ab′) 2 fragment anti-mouse IgG1 (H+L), anti-NOX2 antibody (clone 54.1) and anti-p22 phox antibody (clone 44.1) were from Thermo Fisher Scientific (Courtaboeuf, France).

    Techniques: Derivative Assay, Flow Cytometry, Expressing, Fluorescence, Confocal Microscopy, Staining

    Location of NOX2 in liposome-treated X 0 -CGD iPSC-derived macrophages after C . albicans phagocytosis. Notes: ( A ) Confocal microscopy images showing the staining of NOX2 subunit with 7D5 antibody and AF488-conjugated secondary antibody (green) in X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox or negative liposomes and then for 4 h with C . albicans strain at an MOI of 3:1. ( B ) Z-stack images (top to bottom of cell) of an NOX2/p22 phox liposome-treated X 0 -CGD macrophage incubated for 4 h with C . albicans strain at an MOI of 3:1. Nuclei were counterstained with Hoechst 33258 (blue); scale bars =20 µm in ( A ) and 10 µm in ( B ). The same observations were obtained in at least two experiments. Abbreviations: C. albicans; Candida albicans , CGD, chronic granulomatous disease; iPSC, induced pluripotent stem cell; MOI, multiplicity of infection; X 0 -CGD, X 0 -linked CGD; XCGD, X-linked CGD.

    Journal: International Journal of Nanomedicine

    Article Title: Therapeutic effects of proteoliposomes on X-linked chronic granulomatous disease: proof of concept using macrophages differentiated from patient-specific induced pluripotent stem cells

    doi: 10.2147/IJN.S128611

    Figure Lengend Snippet: Location of NOX2 in liposome-treated X 0 -CGD iPSC-derived macrophages after C . albicans phagocytosis. Notes: ( A ) Confocal microscopy images showing the staining of NOX2 subunit with 7D5 antibody and AF488-conjugated secondary antibody (green) in X 0 -CGD macrophages treated for 8 h with NOX2/p22 phox or negative liposomes and then for 4 h with C . albicans strain at an MOI of 3:1. ( B ) Z-stack images (top to bottom of cell) of an NOX2/p22 phox liposome-treated X 0 -CGD macrophage incubated for 4 h with C . albicans strain at an MOI of 3:1. Nuclei were counterstained with Hoechst 33258 (blue); scale bars =20 µm in ( A ) and 10 µm in ( B ). The same observations were obtained in at least two experiments. Abbreviations: C. albicans; Candida albicans , CGD, chronic granulomatous disease; iPSC, induced pluripotent stem cell; MOI, multiplicity of infection; X 0 -CGD, X 0 -linked CGD; XCGD, X-linked CGD.

    Article Snippet: Paraformaldehyde (PFA), fetal bovine serum (FBS), goat serum, Iscove’s Modified Dulbecco’s Medium (IMDM), Alexa Fluor ® 488-conjugated goat-F(ab′) 2 fragment anti-mouse IgG1 (H+L), anti-NOX2 antibody (clone 54.1) and anti-p22 phox antibody (clone 44.1) were from Thermo Fisher Scientific (Courtaboeuf, France).

    Techniques: Derivative Assay, Confocal Microscopy, Staining, Incubation, Infection

    Analysis of in cellulo toxicity and NADPH oxidase activity restoration in X 0 -CGD iPSC-derived macrophages. Notes: ( A ) Viability of CGDX 0 iPSC-derived macrophages after 8, 12 and 24 h of treatment. Untreated cells are used as positive control (100%) and DMSO-treated cells as negative control. Data are expressed as mean ± SEM ( n =3). The differences of viability between the untreated cells and the liposome-treated cells were analyzed using the nonparametric Mann–Whitney test. ( B ) Morphological images of X 0 -CGD iPSC-derived macrophages after 8, 12 and 24 h of treatment with NOX2/p22 phox liposomes or incubated with IMDM only (magnification ×10). ( C ) X 0 -CGD macrophages were treated for 8 h with NOX2/p22 phox or negative liposomes. Then, WT and X 0 -CGD macrophages were stimulated with PMA (resting; scale bars =50 µm). Abbreviations: CGD, chronic granulomatous disease; DMSO, dimethylsulfoxide; IMDM, Iscove’s Modified Dulbecco’s Medium; iPSC, induced pluripotent stem cell; NADPH, nicotinamide adenine dinucleotide phosphate; ns, nonsignificant; PMA, phorbolmyristate-acetate; SEM, standard error of the mean; WT, wild type; X 0 -CGD, X 0 -linked CGD.

    Journal: International Journal of Nanomedicine

    Article Title: Therapeutic effects of proteoliposomes on X-linked chronic granulomatous disease: proof of concept using macrophages differentiated from patient-specific induced pluripotent stem cells

    doi: 10.2147/IJN.S128611

    Figure Lengend Snippet: Analysis of in cellulo toxicity and NADPH oxidase activity restoration in X 0 -CGD iPSC-derived macrophages. Notes: ( A ) Viability of CGDX 0 iPSC-derived macrophages after 8, 12 and 24 h of treatment. Untreated cells are used as positive control (100%) and DMSO-treated cells as negative control. Data are expressed as mean ± SEM ( n =3). The differences of viability between the untreated cells and the liposome-treated cells were analyzed using the nonparametric Mann–Whitney test. ( B ) Morphological images of X 0 -CGD iPSC-derived macrophages after 8, 12 and 24 h of treatment with NOX2/p22 phox liposomes or incubated with IMDM only (magnification ×10). ( C ) X 0 -CGD macrophages were treated for 8 h with NOX2/p22 phox or negative liposomes. Then, WT and X 0 -CGD macrophages were stimulated with PMA (resting; scale bars =50 µm). Abbreviations: CGD, chronic granulomatous disease; DMSO, dimethylsulfoxide; IMDM, Iscove’s Modified Dulbecco’s Medium; iPSC, induced pluripotent stem cell; NADPH, nicotinamide adenine dinucleotide phosphate; ns, nonsignificant; PMA, phorbolmyristate-acetate; SEM, standard error of the mean; WT, wild type; X 0 -CGD, X 0 -linked CGD.

    Article Snippet: Paraformaldehyde (PFA), fetal bovine serum (FBS), goat serum, Iscove’s Modified Dulbecco’s Medium (IMDM), Alexa Fluor ® 488-conjugated goat-F(ab′) 2 fragment anti-mouse IgG1 (H+L), anti-NOX2 antibody (clone 54.1) and anti-p22 phox antibody (clone 44.1) were from Thermo Fisher Scientific (Courtaboeuf, France).

    Techniques: Activity Assay, Derivative Assay, Positive Control, Negative Control, MANN-WHITNEY, Incubation, Modification