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    Boster†s Phosphate Buffered Saline PBS Powder is a dehydrated raw material reagent intended to be used in the making of prepared media products by reconstitution in distilled water
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    pbs  (Lonza)
    97
    Lonza pbs
    Pbs, supplied by Lonza, used in various techniques. Bioz Stars score: 97/100, based on 1233 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore pbs
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    LI-COR odyssey blocking buffer
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    GE Healthcare pbs
    PLGA <t>NPs</t> containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received <t>PBS.</t> Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p
    Pbs, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 2461 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pbs phosphate buffered saline 10x ph 7 4
    PLGA <t>NPs</t> containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received <t>PBS.</t> Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p
    Pbs Phosphate Buffered Saline 10x Ph 7 4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 291 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad phosphate buffered saline
    PLGA <t>NPs</t> containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received <t>PBS.</t> Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p
    Phosphate Buffered Saline, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1592 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 1x pbs
    PLGA <t>NPs</t> containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received <t>PBS.</t> Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p
    1x Pbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6906 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Beyotime pbs
    PLGA <t>NPs</t> containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received <t>PBS.</t> Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p
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    Fisher Scientific pbs
    PLGA <t>NPs</t> containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received <t>PBS.</t> Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p
    Pbs, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 690 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pbs  (Abcam)
    99
    Abcam pbs
    PLGA <t>NPs</t> containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received <t>PBS.</t> Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p
    Pbs, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 5349 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore phosphate buffer saline pbs
    PLGA <t>NPs</t> containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received <t>PBS.</t> Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p
    Phosphate Buffer Saline Pbs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2095 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson phosphate buffered saline pbs
    Astrogliosis and remyelination after intranasal administration of CNS-targeting Treg. Mice in three groups were given 1 × 10 5 <t>CAR</t> Tregs, CD4 + Mock T cells or <t>PBS</t> alone by intranasal administration at the peak of EAE inflammation (15 days post-EAE immunization). Fifteen days post cell treatment the mice were killed and brain sections from each group (CAR Tregs, Mock CD4 + T cell and PBS) were analyzed for reactive astroglisosis using glial acidic fibrillary protein (GFAP) ( A - I ) and myelination using myelin basic protein (MBP) ( J - R ). GFAP was evaluated in olfactory bulb ( A - C ), corpus callosum ( D - F ), and cerebellum ( G - I ) of brain sagittal section from PBS, Mock CD4 + T cell and CAR Τreg-treated EAE mice. In the olfactory bulb of mice treated with Mock CD4 + T cells there was a strong staining for GFAP whereas this area in PBS-treated mice (A) and of CAR Treg-treated mice (C) exhibited a weak and moderate staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for corpus callosum and cerebellum. MBP was evaluated in brain stem ( J - L ), hippocampus ( M - O ) and cerebellum ( P - R ) of brain sagittal sections in PBS, Mock CD4 + T cell and CAR Treg-treated EAE mice. In the brain stem and cerebellum of mice treated with CAR Tregs, there was a moderate staining for MBP (L, R) whereas the brain stem of Mock-treated mice (K,Q) and PBS-treated mice (J,P) exhibited a weak and strong staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, a moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for hippocampus. Original magnification 10×.
    Phosphate Buffered Saline Pbs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 8489 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phosphate buffered saline
    Astrogliosis and remyelination after intranasal administration of CNS-targeting Treg. Mice in three groups were given 1 × 10 5 <t>CAR</t> Tregs, CD4 + Mock T cells or <t>PBS</t> alone by intranasal administration at the peak of EAE inflammation (15 days post-EAE immunization). Fifteen days post cell treatment the mice were killed and brain sections from each group (CAR Tregs, Mock CD4 + T cell and PBS) were analyzed for reactive astroglisosis using glial acidic fibrillary protein (GFAP) ( A - I ) and myelination using myelin basic protein (MBP) ( J - R ). GFAP was evaluated in olfactory bulb ( A - C ), corpus callosum ( D - F ), and cerebellum ( G - I ) of brain sagittal section from PBS, Mock CD4 + T cell and CAR Τreg-treated EAE mice. In the olfactory bulb of mice treated with Mock CD4 + T cells there was a strong staining for GFAP whereas this area in PBS-treated mice (A) and of CAR Treg-treated mice (C) exhibited a weak and moderate staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for corpus callosum and cerebellum. MBP was evaluated in brain stem ( J - L ), hippocampus ( M - O ) and cerebellum ( P - R ) of brain sagittal sections in PBS, Mock CD4 + T cell and CAR Treg-treated EAE mice. In the brain stem and cerebellum of mice treated with CAR Tregs, there was a moderate staining for MBP (L, R) whereas the brain stem of Mock-treated mice (K,Q) and PBS-treated mice (J,P) exhibited a weak and strong staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, a moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for hippocampus. Original magnification 10×.
    Phosphate Buffered Saline, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1829 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam phosphate buffered saline pbs
    Astrogliosis and remyelination after intranasal administration of CNS-targeting Treg. Mice in three groups were given 1 × 10 5 <t>CAR</t> Tregs, CD4 + Mock T cells or <t>PBS</t> alone by intranasal administration at the peak of EAE inflammation (15 days post-EAE immunization). Fifteen days post cell treatment the mice were killed and brain sections from each group (CAR Tregs, Mock CD4 + T cell and PBS) were analyzed for reactive astroglisosis using glial acidic fibrillary protein (GFAP) ( A - I ) and myelination using myelin basic protein (MBP) ( J - R ). GFAP was evaluated in olfactory bulb ( A - C ), corpus callosum ( D - F ), and cerebellum ( G - I ) of brain sagittal section from PBS, Mock CD4 + T cell and CAR Τreg-treated EAE mice. In the olfactory bulb of mice treated with Mock CD4 + T cells there was a strong staining for GFAP whereas this area in PBS-treated mice (A) and of CAR Treg-treated mice (C) exhibited a weak and moderate staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for corpus callosum and cerebellum. MBP was evaluated in brain stem ( J - L ), hippocampus ( M - O ) and cerebellum ( P - R ) of brain sagittal sections in PBS, Mock CD4 + T cell and CAR Treg-treated EAE mice. In the brain stem and cerebellum of mice treated with CAR Tregs, there was a moderate staining for MBP (L, R) whereas the brain stem of Mock-treated mice (K,Q) and PBS-treated mice (J,P) exhibited a weak and strong staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, a moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for hippocampus. Original magnification 10×.
    Phosphate Buffered Saline Pbs, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 2966 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA pbs
    <t>SPS-neutralization</t> in efficacy tests of SAAP-148 against gram-positive and gram-negative bacteria. Ninety μL of 10 7 CFU/mL MRSA (LUH14616 and Mu50), E. faecalis (ATCC 29212) , P. aeruginosa (PAO1; ATCC BAA47), E. coli (ATCC 35218) and A. baumannii were exposed to 10 μL of 1% (wt/v) SAAP-148 in <t>PBS</t> or PBS for 30 min. Subsequently, samples were homogenized in 500 μL of PBS with or without 0.05% (wt/v) SPS. The means and SD of six independent experiments performed in duplicate are shown. Results are expressed as the number of surviving bacteria in log10 CFU/mL. * indicates significant difference as compared to the samples without SPS (* p
    Pbs, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 92/100, based on 1284 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    PLGA NPs containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received PBS. Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p

    Journal: Vaccines

    Article Title: Adjuvantation of an Influenza Hemagglutinin Antigen with TLR4 and NOD2 Agonists Encapsulated in Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Enhances Immunogenicity and Protection against Lethal Influenza Virus Infection in Mice

    doi: 10.3390/vaccines8030519

    Figure Lengend Snippet: PLGA NPs containing both MPLA and MDP induce stronger antigen-specific CD4+ T-cell response in mice than NPs containing individual PRR agonists. C57BL/6 mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MPLA + MDP with a two-week interval. Non-immunized control mice received PBS. Splenocytes were harvested from mice 14 days after the last immunization. Splenocytes were stained with CFSE, treated with whole hemagglutinin (1 μg/mL). 72 h after cell were harvested for cellular proliferation analysis, cell culture was collected for IFN-γ detection. ( A ) Gating strategy for assessment of proliferating T cells in culture. Single lymphocytes were gated using the forward and side scatter characteristics. Live (DAPI negative) CD3 + T cells were then obtained from the single cells gate. CFSE dim cells were gated in each CD3 + CD4+ as well as CD3 + CD8+ populations. ( B ) % of proliferating CD4 + T-cells (referred to fraction diluted) in response to hemagglutinin restimulation. ( C ) IFN-γ levels in supernatants from HA-stimulated spleen cells, determined by a bio-plex assay 72 h after restimulation. Boxes show interquartile range, whiskers show range, and horizontal lines represent median values. Dots show individual data points. Significant differences ( p

    Article Snippet: PLGA NPs were diluted to 0.1 μg/mL concentration in particle-free 0.02 μm filtered PBS (Whatman, Maidstone, UK).

    Techniques: Mouse Assay, Staining, Cell Culture, Plex Assay

    PLGA NPs containing both MPLA and MDP are more effectively phagocytosed by bone marrow-derived dendritic cells (BMDCs) compared to NPs containing individual PRR agonists. ( A ) A time course of phagocytosis quantification by BMDCs of PLGA NPs containing pHRodo-stained hemagglutinin (HA). Immature BMDCs isolated from naïve C57BL/6 mice were seeded in a 96-well plate at 2 × 10 5 cells per well in complete RPMI medium. The next day, PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MDP-MPLA formulations produced with pHRodo-labeled hemagglutinin were added in a 100 μg/mL final concentration. The medium was removed at the indicated time points, BMDCs were washed with PBS, and the integral fluorescence of each well was determined using a Biotek microplate reader. Each bar represents the mean fold increase ± SD (whiskers) in relative fluorescence units (RFU) over intact (untreated) cells. The mean value of untreated cells (= 1088 ± 10 RFU) was taken equal to 1 (100%). Experiment was performed in triplicate and repeated twice. ( B ) Representative fluorescence microscopy images of BMDCs treated with pHRodo-stained PLGA NPs for 120 min. BMD Cs were stained with anti-CD11c antibodies (green). The cell nucleus was visualized with DAPI (blue). Phagocytosed PLGA NPs were specifically visualized by pHRodo (red) fluorescence. Scale bar represents 20 μm. ( C ) Representative gating strategy used to assess the phagocytic activity of BMDCs. BMDCs were initially gated in an forward versus side scatter (FSC vs. SSC) dot plot and then identified as double positive (MHCII+ CD11c+) population. Phagocytic activity of BMDCs was determined by ( D ) % of pHRodo-positive BMDCs as well as ( E ) fold increase in mean fluorescence intensity (MFI) of total MHCII+ CD11c+ cells relative to untreated cells 120 min after the addition of PLGA NPs. Mean value of untreated cells (= 749.5 ± 9.2 RFU) was taken equal to 1 (100%). Each bar represents the mean ± SD from three independent experiments, each performed in triplicate. Significant differences between PLGA NPs and intact cells are indicated by hashes: ## for p

    Journal: Vaccines

    Article Title: Adjuvantation of an Influenza Hemagglutinin Antigen with TLR4 and NOD2 Agonists Encapsulated in Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Enhances Immunogenicity and Protection against Lethal Influenza Virus Infection in Mice

    doi: 10.3390/vaccines8030519

    Figure Lengend Snippet: PLGA NPs containing both MPLA and MDP are more effectively phagocytosed by bone marrow-derived dendritic cells (BMDCs) compared to NPs containing individual PRR agonists. ( A ) A time course of phagocytosis quantification by BMDCs of PLGA NPs containing pHRodo-stained hemagglutinin (HA). Immature BMDCs isolated from naïve C57BL/6 mice were seeded in a 96-well plate at 2 × 10 5 cells per well in complete RPMI medium. The next day, PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MDP-MPLA formulations produced with pHRodo-labeled hemagglutinin were added in a 100 μg/mL final concentration. The medium was removed at the indicated time points, BMDCs were washed with PBS, and the integral fluorescence of each well was determined using a Biotek microplate reader. Each bar represents the mean fold increase ± SD (whiskers) in relative fluorescence units (RFU) over intact (untreated) cells. The mean value of untreated cells (= 1088 ± 10 RFU) was taken equal to 1 (100%). Experiment was performed in triplicate and repeated twice. ( B ) Representative fluorescence microscopy images of BMDCs treated with pHRodo-stained PLGA NPs for 120 min. BMD Cs were stained with anti-CD11c antibodies (green). The cell nucleus was visualized with DAPI (blue). Phagocytosed PLGA NPs were specifically visualized by pHRodo (red) fluorescence. Scale bar represents 20 μm. ( C ) Representative gating strategy used to assess the phagocytic activity of BMDCs. BMDCs were initially gated in an forward versus side scatter (FSC vs. SSC) dot plot and then identified as double positive (MHCII+ CD11c+) population. Phagocytic activity of BMDCs was determined by ( D ) % of pHRodo-positive BMDCs as well as ( E ) fold increase in mean fluorescence intensity (MFI) of total MHCII+ CD11c+ cells relative to untreated cells 120 min after the addition of PLGA NPs. Mean value of untreated cells (= 749.5 ± 9.2 RFU) was taken equal to 1 (100%). Each bar represents the mean ± SD from three independent experiments, each performed in triplicate. Significant differences between PLGA NPs and intact cells are indicated by hashes: ## for p

    Article Snippet: PLGA NPs were diluted to 0.1 μg/mL concentration in particle-free 0.02 μm filtered PBS (Whatman, Maidstone, UK).

    Techniques: Derivative Assay, Staining, Isolation, Mouse Assay, Produced, Labeling, Concentration Assay, Fluorescence, Microscopy, Activity Assay

    PLGA NPs containing both MPLA and MDP led to a stronger protection in mice than NPs containing individual PRR agonists. Kaplan–Meier survival curves of mice i.m. immunized three times with ( A ) 1 dose (1 mg/mouse, n = 8 per group) or ( B ) 2 doses (2 mg/mouse, n = 10 per group) of PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MDP-MPLA. Non-immunized control mice received PBS. Two weeks after last immunization, mice were infected using a lethal dose (50LD50) of the A/California/07/2009 (H1N1) strain virus. Mortality was monitored daily for 14 days post-challenge. # indicates a significant difference ( p

    Journal: Vaccines

    Article Title: Adjuvantation of an Influenza Hemagglutinin Antigen with TLR4 and NOD2 Agonists Encapsulated in Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Enhances Immunogenicity and Protection against Lethal Influenza Virus Infection in Mice

    doi: 10.3390/vaccines8030519

    Figure Lengend Snippet: PLGA NPs containing both MPLA and MDP led to a stronger protection in mice than NPs containing individual PRR agonists. Kaplan–Meier survival curves of mice i.m. immunized three times with ( A ) 1 dose (1 mg/mouse, n = 8 per group) or ( B ) 2 doses (2 mg/mouse, n = 10 per group) of PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MDP-MPLA. Non-immunized control mice received PBS. Two weeks after last immunization, mice were infected using a lethal dose (50LD50) of the A/California/07/2009 (H1N1) strain virus. Mortality was monitored daily for 14 days post-challenge. # indicates a significant difference ( p

    Article Snippet: PLGA NPs were diluted to 0.1 μg/mL concentration in particle-free 0.02 μm filtered PBS (Whatman, Maidstone, UK).

    Techniques: Mouse Assay, Infection

    PLGA NPs containing both MPLA and MDP lead to a stronger humoral adaptive response in mice than NPs containing individual PRR agonists. BALB/c mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MDP-MPLA. Non-immunized control mice received PBS. Blood was collected from mice 14 days after the last immunization for serum preparation. The geometric mean for 5 mice/group with 95%CI is shown. ( A ) The reciprocal titers of hemagglutinin-specific total IgG antibodies and IgG1, IgG2a, IgG2b, and IgG2c subtypes were detected in serum by ELISA. The geometric mean for 5 mice/group with 95% CI is shown. ( B ) Ratios of IgG2a/IgG1 isotype HA-specific antibodies. ( C ) The reciprocal titers of hemagglutination inhibition (HI) antibodies against the H1N1 virus in the sera of mice immunized with PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MDP-MPLA. Significant differences between PLGA NPs and intact cells are indicated by hashes: # for p

    Journal: Vaccines

    Article Title: Adjuvantation of an Influenza Hemagglutinin Antigen with TLR4 and NOD2 Agonists Encapsulated in Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Enhances Immunogenicity and Protection against Lethal Influenza Virus Infection in Mice

    doi: 10.3390/vaccines8030519

    Figure Lengend Snippet: PLGA NPs containing both MPLA and MDP lead to a stronger humoral adaptive response in mice than NPs containing individual PRR agonists. BALB/c mice ( n = 5/group) were i.m. immunized three times with PLGA NPs: PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MDP-MPLA. Non-immunized control mice received PBS. Blood was collected from mice 14 days after the last immunization for serum preparation. The geometric mean for 5 mice/group with 95%CI is shown. ( A ) The reciprocal titers of hemagglutinin-specific total IgG antibodies and IgG1, IgG2a, IgG2b, and IgG2c subtypes were detected in serum by ELISA. The geometric mean for 5 mice/group with 95% CI is shown. ( B ) Ratios of IgG2a/IgG1 isotype HA-specific antibodies. ( C ) The reciprocal titers of hemagglutination inhibition (HI) antibodies against the H1N1 virus in the sera of mice immunized with PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, and PLGA-HA-MDP-MPLA. Significant differences between PLGA NPs and intact cells are indicated by hashes: # for p

    Article Snippet: PLGA NPs were diluted to 0.1 μg/mL concentration in particle-free 0.02 μm filtered PBS (Whatman, Maidstone, UK).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, HI Assay

    Phagocytosis rate and cell response after the addition of equal amounts of PLGA NPs and H1N1 influenza viral particles to RAW-Blue cells. ( A ) Phagocytosis quantification of PLGA NPs and H1N1 influenza viral particles. RAW-Blue cells were seeded in 96-well plate at 2 × 10 5 cells per well in complete RPMI medium. The next day, normalized to 10 10 particles/well PLGA (roughly equal to 100µg/ml) and virus particles were added to the cells. The medium was removed at the indicated time points, the cells were washed with PBS, and the integral fluorescence (RFU) of each well was determined using a Biotek reader. Mean values (indicated above the bars) of intact cells were taken equal to 1 (100%). Data represent the mean ( n = 3) fold increase over intact cells ± SD. ( B ) NF-κB/AP-1-dependent SEAP reporter gene expression in cell-free culture supernatants from RAW-Blue cells treated for 18 h with PLGA and with virus particles normalized by quantity (10 10 particles/well). Mean values (indicated above the bars) of intact cells were taken equal to 1 (100%). Data represent the mean ( n = 3) fold increase over intact cells ± SD. ( C ) Cytokine levels were measured in cell-free culture supernatants from the same wells using bead-based immunoassay. Bars represent mean fold increase ± SD (whiskers) over intact group. Values (mean concentrations ± SD are indicated above the bars) of intact groups were taken equal to 1 (100%). Significant differences between treated and intact cells are indicated by hashes: # for p

    Journal: Vaccines

    Article Title: Adjuvantation of an Influenza Hemagglutinin Antigen with TLR4 and NOD2 Agonists Encapsulated in Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Enhances Immunogenicity and Protection against Lethal Influenza Virus Infection in Mice

    doi: 10.3390/vaccines8030519

    Figure Lengend Snippet: Phagocytosis rate and cell response after the addition of equal amounts of PLGA NPs and H1N1 influenza viral particles to RAW-Blue cells. ( A ) Phagocytosis quantification of PLGA NPs and H1N1 influenza viral particles. RAW-Blue cells were seeded in 96-well plate at 2 × 10 5 cells per well in complete RPMI medium. The next day, normalized to 10 10 particles/well PLGA (roughly equal to 100µg/ml) and virus particles were added to the cells. The medium was removed at the indicated time points, the cells were washed with PBS, and the integral fluorescence (RFU) of each well was determined using a Biotek reader. Mean values (indicated above the bars) of intact cells were taken equal to 1 (100%). Data represent the mean ( n = 3) fold increase over intact cells ± SD. ( B ) NF-κB/AP-1-dependent SEAP reporter gene expression in cell-free culture supernatants from RAW-Blue cells treated for 18 h with PLGA and with virus particles normalized by quantity (10 10 particles/well). Mean values (indicated above the bars) of intact cells were taken equal to 1 (100%). Data represent the mean ( n = 3) fold increase over intact cells ± SD. ( C ) Cytokine levels were measured in cell-free culture supernatants from the same wells using bead-based immunoassay. Bars represent mean fold increase ± SD (whiskers) over intact group. Values (mean concentrations ± SD are indicated above the bars) of intact groups were taken equal to 1 (100%). Significant differences between treated and intact cells are indicated by hashes: # for p

    Article Snippet: PLGA NPs were diluted to 0.1 μg/mL concentration in particle-free 0.02 μm filtered PBS (Whatman, Maidstone, UK).

    Techniques: Fluorescence, Expressing, Bead-based Assay

    PLGA NPs containing both MPLA and MDP induce stronger NF-κB transcriptional response in BALB/c-Tg (Rela-luc) 31Xen transgenic mice than NPs with individual PRR agonists. ( A ) Representative bioluminescent pseudocolored images of NF-κB-luciferase activity in live NF-κB-Luc transgenic mice 3 h after i.m. injection of PBS, PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, or PLGA-HA-MPLA + MDP (all in 1 mg/mouse in 100 μL). Prior to imaging, mice received an i.p. injection of luciferin followed by anesthesia. Non-immunized control mice received PBS. The intensity of bioluminescence (i.e., NF- κB activity) is shown in color according to the scale on the right. Two additional mice in each group gave similar results. ( B ) NF-κB-dependent bioluminescence in tissue homogenates prepared from NF-κB-Luc transgenic mice at the same time point after the injection of PLGA NPs. Control mice were injected with PBS. Values of control group (mean fluorescence intensities ± SD are indicated above the bars) were taken equal to 1 (100%). Results are expressed as the fold increase in relative luminescence units over PBS-treated control animals. Each bar represents the mean of three mice per group ± SD (error bars). Significant differences between PLGA NPs and PBS treated mice are indicated by hashes: ## for p

    Journal: Vaccines

    Article Title: Adjuvantation of an Influenza Hemagglutinin Antigen with TLR4 and NOD2 Agonists Encapsulated in Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Enhances Immunogenicity and Protection against Lethal Influenza Virus Infection in Mice

    doi: 10.3390/vaccines8030519

    Figure Lengend Snippet: PLGA NPs containing both MPLA and MDP induce stronger NF-κB transcriptional response in BALB/c-Tg (Rela-luc) 31Xen transgenic mice than NPs with individual PRR agonists. ( A ) Representative bioluminescent pseudocolored images of NF-κB-luciferase activity in live NF-κB-Luc transgenic mice 3 h after i.m. injection of PBS, PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, or PLGA-HA-MPLA + MDP (all in 1 mg/mouse in 100 μL). Prior to imaging, mice received an i.p. injection of luciferin followed by anesthesia. Non-immunized control mice received PBS. The intensity of bioluminescence (i.e., NF- κB activity) is shown in color according to the scale on the right. Two additional mice in each group gave similar results. ( B ) NF-κB-dependent bioluminescence in tissue homogenates prepared from NF-κB-Luc transgenic mice at the same time point after the injection of PLGA NPs. Control mice were injected with PBS. Values of control group (mean fluorescence intensities ± SD are indicated above the bars) were taken equal to 1 (100%). Results are expressed as the fold increase in relative luminescence units over PBS-treated control animals. Each bar represents the mean of three mice per group ± SD (error bars). Significant differences between PLGA NPs and PBS treated mice are indicated by hashes: ## for p

    Article Snippet: PLGA NPs were diluted to 0.1 μg/mL concentration in particle-free 0.02 μm filtered PBS (Whatman, Maidstone, UK).

    Techniques: Transgenic Assay, Mouse Assay, Luciferase, Activity Assay, Injection, Imaging, Fluorescence

    PLGA NPs containing both MPLA and MDP induce a stronger local cytokine response in mice than NPs containing individual PRR agonists. BALB/c mice were intramuscularly injected with 1 mg/dose of PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, or PLGA-HA-MPLA + MDP. Non-immunized control mice received PBS. Three hours later, inguinal lymph nodes, the liver, the spleen, and the thigh muscle (corresponding to the injection site) were isolated. Cytokine levels were measured in tissue homogenates using bead-based immunoassay. Control mice were injected with PBS. Results are expressed as the mean fold change in cytokine concentration relative to the mean concentration in PBS-treated control animals. Values of control group (mean concentrations ± SD are indicated above the bars) were taken equal to 1 (100%). Each point represents the mean of three mice per group ± SD (error bars). Significant differences between PLGA NPs and intact cells are indicated by hashes: # for p

    Journal: Vaccines

    Article Title: Adjuvantation of an Influenza Hemagglutinin Antigen with TLR4 and NOD2 Agonists Encapsulated in Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Enhances Immunogenicity and Protection against Lethal Influenza Virus Infection in Mice

    doi: 10.3390/vaccines8030519

    Figure Lengend Snippet: PLGA NPs containing both MPLA and MDP induce a stronger local cytokine response in mice than NPs containing individual PRR agonists. BALB/c mice were intramuscularly injected with 1 mg/dose of PLGA-HA, PLGA-HA-MPLA, PLGA-HA-MDP, or PLGA-HA-MPLA + MDP. Non-immunized control mice received PBS. Three hours later, inguinal lymph nodes, the liver, the spleen, and the thigh muscle (corresponding to the injection site) were isolated. Cytokine levels were measured in tissue homogenates using bead-based immunoassay. Control mice were injected with PBS. Results are expressed as the mean fold change in cytokine concentration relative to the mean concentration in PBS-treated control animals. Values of control group (mean concentrations ± SD are indicated above the bars) were taken equal to 1 (100%). Each point represents the mean of three mice per group ± SD (error bars). Significant differences between PLGA NPs and intact cells are indicated by hashes: # for p

    Article Snippet: PLGA NPs were diluted to 0.1 μg/mL concentration in particle-free 0.02 μm filtered PBS (Whatman, Maidstone, UK).

    Techniques: Mouse Assay, Injection, Isolation, Bead-based Assay, Concentration Assay

    Astrogliosis and remyelination after intranasal administration of CNS-targeting Treg. Mice in three groups were given 1 × 10 5 CAR Tregs, CD4 + Mock T cells or PBS alone by intranasal administration at the peak of EAE inflammation (15 days post-EAE immunization). Fifteen days post cell treatment the mice were killed and brain sections from each group (CAR Tregs, Mock CD4 + T cell and PBS) were analyzed for reactive astroglisosis using glial acidic fibrillary protein (GFAP) ( A - I ) and myelination using myelin basic protein (MBP) ( J - R ). GFAP was evaluated in olfactory bulb ( A - C ), corpus callosum ( D - F ), and cerebellum ( G - I ) of brain sagittal section from PBS, Mock CD4 + T cell and CAR Τreg-treated EAE mice. In the olfactory bulb of mice treated with Mock CD4 + T cells there was a strong staining for GFAP whereas this area in PBS-treated mice (A) and of CAR Treg-treated mice (C) exhibited a weak and moderate staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for corpus callosum and cerebellum. MBP was evaluated in brain stem ( J - L ), hippocampus ( M - O ) and cerebellum ( P - R ) of brain sagittal sections in PBS, Mock CD4 + T cell and CAR Treg-treated EAE mice. In the brain stem and cerebellum of mice treated with CAR Tregs, there was a moderate staining for MBP (L, R) whereas the brain stem of Mock-treated mice (K,Q) and PBS-treated mice (J,P) exhibited a weak and strong staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, a moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for hippocampus. Original magnification 10×.

    Journal: Journal of Neuroinflammation

    Article Title: CAR/FoxP3-engineered T regulatory cells target the CNS and suppress EAE upon intranasal delivery

    doi: 10.1186/1742-2094-9-112

    Figure Lengend Snippet: Astrogliosis and remyelination after intranasal administration of CNS-targeting Treg. Mice in three groups were given 1 × 10 5 CAR Tregs, CD4 + Mock T cells or PBS alone by intranasal administration at the peak of EAE inflammation (15 days post-EAE immunization). Fifteen days post cell treatment the mice were killed and brain sections from each group (CAR Tregs, Mock CD4 + T cell and PBS) were analyzed for reactive astroglisosis using glial acidic fibrillary protein (GFAP) ( A - I ) and myelination using myelin basic protein (MBP) ( J - R ). GFAP was evaluated in olfactory bulb ( A - C ), corpus callosum ( D - F ), and cerebellum ( G - I ) of brain sagittal section from PBS, Mock CD4 + T cell and CAR Τreg-treated EAE mice. In the olfactory bulb of mice treated with Mock CD4 + T cells there was a strong staining for GFAP whereas this area in PBS-treated mice (A) and of CAR Treg-treated mice (C) exhibited a weak and moderate staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for corpus callosum and cerebellum. MBP was evaluated in brain stem ( J - L ), hippocampus ( M - O ) and cerebellum ( P - R ) of brain sagittal sections in PBS, Mock CD4 + T cell and CAR Treg-treated EAE mice. In the brain stem and cerebellum of mice treated with CAR Tregs, there was a moderate staining for MBP (L, R) whereas the brain stem of Mock-treated mice (K,Q) and PBS-treated mice (J,P) exhibited a weak and strong staining, respectively. There is an extremely weak staining in PBS-treated EAE mice, a moderate staining in Mock CD4 + T cell-treated EAE mice and strong staining in CAR Treg-treated EAE mice for hippocampus. Original magnification 10×.

    Article Snippet: Cells (1 × 105 CAR or Mock-transduced Tregs diluted in 10uL PBS) or phosphate-buffered saline (PBS) were administered i.n. in 5μL PBS using a plastic catheter connected to a pipette (polyethylene tube, Becton Dickinson, Franklin Lakes, NJ, USA) inserted for 3 mm in both nasal nostrils during anesthesia (0.05 to 0.1 mg ketalamine-xylazine mixture/10 g body weight; ketamine 50 mg/ml, Pfizer AB, Sollentuna, Sweden; xylazine 20 mg/ml, Bayer AG Animal Health, Business Group, Leverkusen, Germany).

    Techniques: Mouse Assay, Staining

    Engineered Tregs localize to the CNS when administered intranasally in naive mice. Treg cells were administered intranasally in the right nostril and the distribution of green fluorescent protein (GFP) in horizontal cryosections of the brain of naïve mice was studied 24 hours after the delivery. The schematic drawing describes a selective immunofluorescence in various brain regions (green spots). GFP immunofluorescence is present in the granular and to a lower extent in the external plexiform layer of the olfactory bulb (B, C) , lateral septal nucleus (E) , central medial thalamic nucleus (F) , ectorhinal cortex (H) , medial genic nucleus (I) and cerebellum (K, L) of a CAR Treg-treated naïve mouse. Corresponding areas showing no GFP fluorescence in a PBS-treated naïve mouse are (A, D, G, J) . Enlargements of areas in olfactory bulb and cerebellum (as indicated by boxes) are depicted in C and L. Detail of GFP immunofluorescence in the central medial thalamic nucleus and medial genic nucleus (F, I). Cell nuclei (blue) are stained with DAPI Original magnification 10× (A, B, D, E, G, H, J, K) and 40× (C, F, I, L).

    Journal: Journal of Neuroinflammation

    Article Title: CAR/FoxP3-engineered T regulatory cells target the CNS and suppress EAE upon intranasal delivery

    doi: 10.1186/1742-2094-9-112

    Figure Lengend Snippet: Engineered Tregs localize to the CNS when administered intranasally in naive mice. Treg cells were administered intranasally in the right nostril and the distribution of green fluorescent protein (GFP) in horizontal cryosections of the brain of naïve mice was studied 24 hours after the delivery. The schematic drawing describes a selective immunofluorescence in various brain regions (green spots). GFP immunofluorescence is present in the granular and to a lower extent in the external plexiform layer of the olfactory bulb (B, C) , lateral septal nucleus (E) , central medial thalamic nucleus (F) , ectorhinal cortex (H) , medial genic nucleus (I) and cerebellum (K, L) of a CAR Treg-treated naïve mouse. Corresponding areas showing no GFP fluorescence in a PBS-treated naïve mouse are (A, D, G, J) . Enlargements of areas in olfactory bulb and cerebellum (as indicated by boxes) are depicted in C and L. Detail of GFP immunofluorescence in the central medial thalamic nucleus and medial genic nucleus (F, I). Cell nuclei (blue) are stained with DAPI Original magnification 10× (A, B, D, E, G, H, J, K) and 40× (C, F, I, L).

    Article Snippet: Cells (1 × 105 CAR or Mock-transduced Tregs diluted in 10uL PBS) or phosphate-buffered saline (PBS) were administered i.n. in 5μL PBS using a plastic catheter connected to a pipette (polyethylene tube, Becton Dickinson, Franklin Lakes, NJ, USA) inserted for 3 mm in both nasal nostrils during anesthesia (0.05 to 0.1 mg ketalamine-xylazine mixture/10 g body weight; ketamine 50 mg/ml, Pfizer AB, Sollentuna, Sweden; xylazine 20 mg/ml, Bayer AG Animal Health, Business Group, Leverkusen, Germany).

    Techniques: Mouse Assay, Immunofluorescence, Fluorescence, Staining

    CNS-targeting Tregs can reduce EAE symptoms. ( A ) Ten mice in three groups were given 1 × 10 5 CAR Tregs, CD4 + Mock T cells or PBS alone by i.n. administration at the peak of EAE inflammation (15 days post-EAE immunization) and thereafter were monitored for EAE symptoms. Ten days post cell treatment all EAE mice in the CAR Treg group were cured ( P

    Journal: Journal of Neuroinflammation

    Article Title: CAR/FoxP3-engineered T regulatory cells target the CNS and suppress EAE upon intranasal delivery

    doi: 10.1186/1742-2094-9-112

    Figure Lengend Snippet: CNS-targeting Tregs can reduce EAE symptoms. ( A ) Ten mice in three groups were given 1 × 10 5 CAR Tregs, CD4 + Mock T cells or PBS alone by i.n. administration at the peak of EAE inflammation (15 days post-EAE immunization) and thereafter were monitored for EAE symptoms. Ten days post cell treatment all EAE mice in the CAR Treg group were cured ( P

    Article Snippet: Cells (1 × 105 CAR or Mock-transduced Tregs diluted in 10uL PBS) or phosphate-buffered saline (PBS) were administered i.n. in 5μL PBS using a plastic catheter connected to a pipette (polyethylene tube, Becton Dickinson, Franklin Lakes, NJ, USA) inserted for 3 mm in both nasal nostrils during anesthesia (0.05 to 0.1 mg ketalamine-xylazine mixture/10 g body weight; ketamine 50 mg/ml, Pfizer AB, Sollentuna, Sweden; xylazine 20 mg/ml, Bayer AG Animal Health, Business Group, Leverkusen, Germany).

    Techniques: Mouse Assay

    Decreased expression of effector cytokines in CNS-targeting CAR Treg-treated brain. Mice in three groups were given 1 × 10 5 CAR Tregs, CD4 + Mock T cells or PBS alone by i.n. administration at the peak of EAE inflammation (15 days post-EAE immunization). Fifteen days post cell treatment, brain biopsies from five EAE mice per group (CAR Tregs, Mock CD4 + T cells and PBS) were analyzed for expression of effector cytokines (IL-12 and IFNγ) by quantitative RT-PCR. Error bars represent standard error of the mean (SEM).

    Journal: Journal of Neuroinflammation

    Article Title: CAR/FoxP3-engineered T regulatory cells target the CNS and suppress EAE upon intranasal delivery

    doi: 10.1186/1742-2094-9-112

    Figure Lengend Snippet: Decreased expression of effector cytokines in CNS-targeting CAR Treg-treated brain. Mice in three groups were given 1 × 10 5 CAR Tregs, CD4 + Mock T cells or PBS alone by i.n. administration at the peak of EAE inflammation (15 days post-EAE immunization). Fifteen days post cell treatment, brain biopsies from five EAE mice per group (CAR Tregs, Mock CD4 + T cells and PBS) were analyzed for expression of effector cytokines (IL-12 and IFNγ) by quantitative RT-PCR. Error bars represent standard error of the mean (SEM).

    Article Snippet: Cells (1 × 105 CAR or Mock-transduced Tregs diluted in 10uL PBS) or phosphate-buffered saline (PBS) were administered i.n. in 5μL PBS using a plastic catheter connected to a pipette (polyethylene tube, Becton Dickinson, Franklin Lakes, NJ, USA) inserted for 3 mm in both nasal nostrils during anesthesia (0.05 to 0.1 mg ketalamine-xylazine mixture/10 g body weight; ketamine 50 mg/ml, Pfizer AB, Sollentuna, Sweden; xylazine 20 mg/ml, Bayer AG Animal Health, Business Group, Leverkusen, Germany).

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR

    SPS-neutralization in efficacy tests of SAAP-148 against gram-positive and gram-negative bacteria. Ninety μL of 10 7 CFU/mL MRSA (LUH14616 and Mu50), E. faecalis (ATCC 29212) , P. aeruginosa (PAO1; ATCC BAA47), E. coli (ATCC 35218) and A. baumannii were exposed to 10 μL of 1% (wt/v) SAAP-148 in PBS or PBS for 30 min. Subsequently, samples were homogenized in 500 μL of PBS with or without 0.05% (wt/v) SPS. The means and SD of six independent experiments performed in duplicate are shown. Results are expressed as the number of surviving bacteria in log10 CFU/mL. * indicates significant difference as compared to the samples without SPS (* p

    Journal: BMC Infectious Diseases

    Article Title: SPS-neutralization in tissue samples for efficacy testing of antimicrobial peptides

    doi: 10.1186/s12879-019-4700-1

    Figure Lengend Snippet: SPS-neutralization in efficacy tests of SAAP-148 against gram-positive and gram-negative bacteria. Ninety μL of 10 7 CFU/mL MRSA (LUH14616 and Mu50), E. faecalis (ATCC 29212) , P. aeruginosa (PAO1; ATCC BAA47), E. coli (ATCC 35218) and A. baumannii were exposed to 10 μL of 1% (wt/v) SAAP-148 in PBS or PBS for 30 min. Subsequently, samples were homogenized in 500 μL of PBS with or without 0.05% (wt/v) SPS. The means and SD of six independent experiments performed in duplicate are shown. Results are expressed as the number of surviving bacteria in log10 CFU/mL. * indicates significant difference as compared to the samples without SPS (* p

    Article Snippet: SPS-neutralization of antimicrobial activity Ten μL of PBS or one of the antimicrobial agents: 1% (wt/v) SAAP-148 in PBS, 1% (wt/v) pexiganan in PBS, 1% (wt/wt) SSD, 0.5% (v/v) chlorhexidine in 70% alcohol, 2% (wt/wt) Bactroban or 2% (wt/wt) Fucidin were added to polypropylene vials containing 400 μL of PBS, 0.05, 0.1, 0.5% or 1% (wt/v; final concentrations) SPS (Fig. ) in PBS (Merck, KGaA, Darmstadt, Germany).

    Techniques: Neutralization

    Effect of SPS on the antimicrobial activity of various antimicrobial agents. Mixtures of 10 μL of SAAP-148 (1% wt/v), pexiganan (1% wt/v), chlorhexidine (0.5% v/v in 70% alcohol) or PBS and 400 μL of PBS, 0.05, 0.1, 0.5% or 1% (wt/v; final concentrations) SPS in PBS were prepared. Ninety μL of LUH14616 with a final concentration of 10 5 CFU/mL were added to these mixtures to determine the antimicrobial activity after 30 min incubation at 37 °C and 5% CO 2 . The means and standard deviations (SD) of three independent experiments performed in duplicate are shown. Results are expressed as the number of surviving bacteria in log10 CFU/mL. * indicates significant difference as compared to the samples without SPS (* p

    Journal: BMC Infectious Diseases

    Article Title: SPS-neutralization in tissue samples for efficacy testing of antimicrobial peptides

    doi: 10.1186/s12879-019-4700-1

    Figure Lengend Snippet: Effect of SPS on the antimicrobial activity of various antimicrobial agents. Mixtures of 10 μL of SAAP-148 (1% wt/v), pexiganan (1% wt/v), chlorhexidine (0.5% v/v in 70% alcohol) or PBS and 400 μL of PBS, 0.05, 0.1, 0.5% or 1% (wt/v; final concentrations) SPS in PBS were prepared. Ninety μL of LUH14616 with a final concentration of 10 5 CFU/mL were added to these mixtures to determine the antimicrobial activity after 30 min incubation at 37 °C and 5% CO 2 . The means and standard deviations (SD) of three independent experiments performed in duplicate are shown. Results are expressed as the number of surviving bacteria in log10 CFU/mL. * indicates significant difference as compared to the samples without SPS (* p

    Article Snippet: SPS-neutralization of antimicrobial activity Ten μL of PBS or one of the antimicrobial agents: 1% (wt/v) SAAP-148 in PBS, 1% (wt/v) pexiganan in PBS, 1% (wt/wt) SSD, 0.5% (v/v) chlorhexidine in 70% alcohol, 2% (wt/wt) Bactroban or 2% (wt/wt) Fucidin were added to polypropylene vials containing 400 μL of PBS, 0.05, 0.1, 0.5% or 1% (wt/v; final concentrations) SPS (Fig. ) in PBS (Merck, KGaA, Darmstadt, Germany).

    Techniques: Activity Assay, Concentration Assay, Incubation

    SPS-neutralization of residual activity of various antimicrobial agents. Excision wound models were inoculated with 10 5 CFU/mL LUH14616 for 1 h and exposed to 20 μL of SAAP-148 (1% wt/v), pexiganan (1% wt/v), chlorhexidine (0.5% v/v in 70% alcohol) or PBS for 1 h. Subsequently, the models were homogenized in 1 mL of PBS with or without 0.05% (wt/v) SPS. The means and SD of at least eight independent experiments performed in triplicate are shown. Results are expressed as the number of surviving bacteria in log10 CFU/mL. * indicates significant difference as compared to the samples without SPS (* p

    Journal: BMC Infectious Diseases

    Article Title: SPS-neutralization in tissue samples for efficacy testing of antimicrobial peptides

    doi: 10.1186/s12879-019-4700-1

    Figure Lengend Snippet: SPS-neutralization of residual activity of various antimicrobial agents. Excision wound models were inoculated with 10 5 CFU/mL LUH14616 for 1 h and exposed to 20 μL of SAAP-148 (1% wt/v), pexiganan (1% wt/v), chlorhexidine (0.5% v/v in 70% alcohol) or PBS for 1 h. Subsequently, the models were homogenized in 1 mL of PBS with or without 0.05% (wt/v) SPS. The means and SD of at least eight independent experiments performed in triplicate are shown. Results are expressed as the number of surviving bacteria in log10 CFU/mL. * indicates significant difference as compared to the samples without SPS (* p

    Article Snippet: SPS-neutralization of antimicrobial activity Ten μL of PBS or one of the antimicrobial agents: 1% (wt/v) SAAP-148 in PBS, 1% (wt/v) pexiganan in PBS, 1% (wt/wt) SSD, 0.5% (v/v) chlorhexidine in 70% alcohol, 2% (wt/wt) Bactroban or 2% (wt/wt) Fucidin were added to polypropylene vials containing 400 μL of PBS, 0.05, 0.1, 0.5% or 1% (wt/v; final concentrations) SPS (Fig. ) in PBS (Merck, KGaA, Darmstadt, Germany).

    Techniques: Neutralization, Activity Assay