alexa fluor 488 conjugated goat anti mouse igg antibody  (Thermo Fisher)


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    Name:
    F ab 2 Goat anti Mouse IgG IgM H L Secondary Antibody
    Description:
    F ab 2 Goat anti Mouse IgG IgM H L Secondary Antibody for IF Flow
    Catalog Number:
    A10683
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Cell Analysis|Secondary Detection
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    Structured Review

    Thermo Fisher alexa fluor 488 conjugated goat anti mouse igg antibody
    Cortical punctate structures that contain RCNMV MP are sites of viral RNA replication. Nicotiana benthamiana protoplasts were inoculated with recombinant RCNMV RNAs that expressed the MP-mCherry fusion protein ( Figure S1B ) and subjected to immunostaining with anti-dsRNA primary antibody followed by <t>Alexa</t> Fluor 488-conjugated secondary antibody at an early stage of infection (16 hpi, left 2 rows of panels) and at a late stage of infection (24 hpi, center 2 rows of panels). The right-most panels show the results for mock-inoculated protoplasts treated with the same antibodies. Images present confocal projections of five optical sections at 1 µm intervals, which range from the surface to the middle of the protoplasts. DIC: differential interference contrast, N: nucleus. Scale bar = 20 µm.
    F ab 2 Goat anti Mouse IgG IgM H L Secondary Antibody for IF Flow
    https://www.bioz.com/result/alexa fluor 488 conjugated goat anti mouse igg antibody/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alexa fluor 488 conjugated goat anti mouse igg antibody - by Bioz Stars, 2021-05
    86/100 stars

    Images

    1) Product Images from "GAPDH-A Recruits a Plant Virus Movement Protein to Cortical Virus Replication Complexes to Facilitate Viral Cell-to-Cell Movement"

    Article Title: GAPDH-A Recruits a Plant Virus Movement Protein to Cortical Virus Replication Complexes to Facilitate Viral Cell-to-Cell Movement

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1004505

    Cortical punctate structures that contain RCNMV MP are sites of viral RNA replication. Nicotiana benthamiana protoplasts were inoculated with recombinant RCNMV RNAs that expressed the MP-mCherry fusion protein ( Figure S1B ) and subjected to immunostaining with anti-dsRNA primary antibody followed by Alexa Fluor 488-conjugated secondary antibody at an early stage of infection (16 hpi, left 2 rows of panels) and at a late stage of infection (24 hpi, center 2 rows of panels). The right-most panels show the results for mock-inoculated protoplasts treated with the same antibodies. Images present confocal projections of five optical sections at 1 µm intervals, which range from the surface to the middle of the protoplasts. DIC: differential interference contrast, N: nucleus. Scale bar = 20 µm.
    Figure Legend Snippet: Cortical punctate structures that contain RCNMV MP are sites of viral RNA replication. Nicotiana benthamiana protoplasts were inoculated with recombinant RCNMV RNAs that expressed the MP-mCherry fusion protein ( Figure S1B ) and subjected to immunostaining with anti-dsRNA primary antibody followed by Alexa Fluor 488-conjugated secondary antibody at an early stage of infection (16 hpi, left 2 rows of panels) and at a late stage of infection (24 hpi, center 2 rows of panels). The right-most panels show the results for mock-inoculated protoplasts treated with the same antibodies. Images present confocal projections of five optical sections at 1 µm intervals, which range from the surface to the middle of the protoplasts. DIC: differential interference contrast, N: nucleus. Scale bar = 20 µm.

    Techniques Used: Recombinant, Immunostaining, Infection

    Related Articles

    Cell Culture:

    Article Title: IL-10-producing regulatory B cells induced by IL-33 (BregIL-33) effectively attenuate mucosal inflammatory responses in the gut
    Article Snippet: 4.7 In vitro BregIL-33 suppression assays The in vitro suppressive capacity of BregIL-33 on B responder cell proliferation and division was determined by the 3 H-thymidine uptake and CFSE dilution assays. .. Respectively, fixed numbers (105 ) of B responder cells (CD19+ CD23+ ) were cultured, in the presence or absence of anti-CD40 (2.5 μg/ml, Enzo Life Sciences, UK), anti-IgM (5 μg/ml, Thermo Scientific, UK) or LPS (0.5 μg/ml, Sigma–Aldrich, UK), with titrated doses of BregIL-33 (CD19+ CD23− ) isolated from IL-33-treated WT or IL-10−/− mice as described above. .. In some experiments, purified BregIL-33 were first primed by the anti-CD40 antibody (2.5 μg/ml, 5 h) before adding them to the responder cells.

    Article Title: Immune Tolerance to Apoptotic Self Is Mediated Primarily by Regulatory B1a Cells
    Article Snippet: On D7 mice were sacrificed and blood, peritoneal lavage and spleens harvested. .. Splenic CD19+ve B cells were FACs sorted into IL-10-GFP+ve or −ve fractions, phenotyped and cultured for 10 days in the presence of MegaAPRIL [Adipogen (200 ng/ml)], CpGB [ODN1826 Eurofins MWG Operon (1 μg/ml)], and IL-4 [R & D System (50 ng/ml)], after which culture supernatants were checked for IgM and IgG levels (Ready-SET-Go ELISA kit eBioScience). .. Hybridoma Generation Highly purified IL-10-GFP+ve B1a cells, generated in vivo , were fused with SP2/0 cells to generate hybridomas.

    Isolation:

    Article Title: IL-10-producing regulatory B cells induced by IL-33 (BregIL-33) effectively attenuate mucosal inflammatory responses in the gut
    Article Snippet: 4.7 In vitro BregIL-33 suppression assays The in vitro suppressive capacity of BregIL-33 on B responder cell proliferation and division was determined by the 3 H-thymidine uptake and CFSE dilution assays. .. Respectively, fixed numbers (105 ) of B responder cells (CD19+ CD23+ ) were cultured, in the presence or absence of anti-CD40 (2.5 μg/ml, Enzo Life Sciences, UK), anti-IgM (5 μg/ml, Thermo Scientific, UK) or LPS (0.5 μg/ml, Sigma–Aldrich, UK), with titrated doses of BregIL-33 (CD19+ CD23− ) isolated from IL-33-treated WT or IL-10−/− mice as described above. .. In some experiments, purified BregIL-33 were first primed by the anti-CD40 antibody (2.5 μg/ml, 5 h) before adding them to the responder cells.

    Mouse Assay:

    Article Title: IL-10-producing regulatory B cells induced by IL-33 (BregIL-33) effectively attenuate mucosal inflammatory responses in the gut
    Article Snippet: 4.7 In vitro BregIL-33 suppression assays The in vitro suppressive capacity of BregIL-33 on B responder cell proliferation and division was determined by the 3 H-thymidine uptake and CFSE dilution assays. .. Respectively, fixed numbers (105 ) of B responder cells (CD19+ CD23+ ) were cultured, in the presence or absence of anti-CD40 (2.5 μg/ml, Enzo Life Sciences, UK), anti-IgM (5 μg/ml, Thermo Scientific, UK) or LPS (0.5 μg/ml, Sigma–Aldrich, UK), with titrated doses of BregIL-33 (CD19+ CD23− ) isolated from IL-33-treated WT or IL-10−/− mice as described above. .. In some experiments, purified BregIL-33 were first primed by the anti-CD40 antibody (2.5 μg/ml, 5 h) before adding them to the responder cells.

    Incubation:

    Article Title: Cryptococcus neoformans induces antimicrobial responses and behaves as a facultative intracellular pathogen in the non mammalian model Galleria mellonella
    Article Snippet: The cells were incubated for 30 min at 37°C, and washed with PBS/BSA. .. In the case of samples incubated mAb 2H1 and 2D10, a goat anti-mouse (GAM) IgG-Alexa 488 (Life Technologies, Reference A11001) or GAM IgM secondary Ab conjugated to Alexa 488 (Life Technologies, Reference ) was added at 1 μg/mL, respectively. ..

    Article Title: Secreted NS1 of Dengue Virus Attaches to the Surface of Cells via Interactions with Heparan Sulfate and Chondroitin Sulfate E
    Article Snippet: A mixture of IgG1 (MOPC-21, Sigma) and IgG2a (UPC-10, Sigma) at 20 μg/ml was used as isotype control Abs. .. After three washes, sections were incubated with Alexa Fluor 488 conjugated goat anti-mouse IgG secondary antibody (Invitrogen) and followed by 1-h incubation at room temperature with rabbit anti-human CD31 (PECAM) (Santa Cruz Biotechnology) at the dilution of 1:10. .. Sections were washed three times and incubated with Cy3 conjugated donkey anti-rabbit IgG (Jackson Immuno Research Laboratories).

    other:

    Article Title: Expansion of polyreactive B cells cross-reactive to HLA and self in the blood of a patient with kidney graft rejection
    Article Snippet: Antibodies reactive to beads were detected with an anti-IgG (One Lambda) or IgM (Invitrogen, Carlsbad, CA) PE-conjugated secondary antibody on a Luminex 200 apparatus (Luminex, Austin, TX).

    FACS:

    Article Title: Immune Tolerance to Apoptotic Self Is Mediated Primarily by Regulatory B1a Cells
    Article Snippet: On D7 mice were sacrificed and blood, peritoneal lavage and spleens harvested. .. Splenic CD19+ve B cells were FACs sorted into IL-10-GFP+ve or −ve fractions, phenotyped and cultured for 10 days in the presence of MegaAPRIL [Adipogen (200 ng/ml)], CpGB [ODN1826 Eurofins MWG Operon (1 μg/ml)], and IL-4 [R & D System (50 ng/ml)], after which culture supernatants were checked for IgM and IgG levels (Ready-SET-Go ELISA kit eBioScience). .. Hybridoma Generation Highly purified IL-10-GFP+ve B1a cells, generated in vivo , were fused with SP2/0 cells to generate hybridomas.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Immune Tolerance to Apoptotic Self Is Mediated Primarily by Regulatory B1a Cells
    Article Snippet: On D7 mice were sacrificed and blood, peritoneal lavage and spleens harvested. .. Splenic CD19+ve B cells were FACs sorted into IL-10-GFP+ve or −ve fractions, phenotyped and cultured for 10 days in the presence of MegaAPRIL [Adipogen (200 ng/ml)], CpGB [ODN1826 Eurofins MWG Operon (1 μg/ml)], and IL-4 [R & D System (50 ng/ml)], after which culture supernatants were checked for IgM and IgG levels (Ready-SET-Go ELISA kit eBioScience). .. Hybridoma Generation Highly purified IL-10-GFP+ve B1a cells, generated in vivo , were fused with SP2/0 cells to generate hybridomas.

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    Thermo Fisher alexa fluor 488 conjugated goat anti mouse igg antibody
    Chemical structures of the compounds tested and visualization of SARS-CoV-2-infected Vero cells. ( A ) Chemical structures of gemcitabine, 2FdC, and remdesivir. ( B ) MTT-based cytotoxicity assay of gemcitabine at different concentrations and time points. Vero cells were treated with increasing concentrations of gemcitabine for 24 h (black square) or 48 h (gray square). Percentage cell viability was measured by using MTT, in which mock-treated cells served as a control (100%). ( C ) Fluorescein diacetate-based cytotoxicity assay of gemcitabine. Vero cells were treated with increasing concentrations of gemcitabine for 24 (black square) and 48 h (gray square). Percentage cell viability was measured by addition of fluorescein diacetate, in which mock-treated cells served as a control (100%). ( D ) Fluorescein diacetate-based cytotoxicity assay of a delivery vehicle. Increasing concentrations of DMSO, 0.2, 0.6 and 1.8% ( v / v ), that were identically included in 100, 300 and 900 μM gemcitabine shown in ( C ), were treated to Vero cells for 24 (black bar) and 48 h (gray bar). Values in ( B – D ) are means ± standard deviations from thee three independent experiments. ( E ) Visualization of SARS-CoV-2 infection. Vero cells were mock-infected (Mock) or infected with SARS-CoV-2 at a multiplicity of infection (MOI) of 0.02 for 24 h. Viral spike (S) protein was probed with mouse anti-S antibody and <t>Alexa</t> Fluor 488-conjugated goat anti-mouse antibody (green). Cellular nuclei were counterstained with DAPI (blue). Magnification ×20.
    Alexa Fluor 488 Conjugated Goat Anti Mouse Igg 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/alexa fluor 488 conjugated goat anti mouse igg antibody/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alexa fluor 488 conjugated goat anti mouse igg antibody - by Bioz Stars, 2021-05
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    99
    Thermo Fisher alexa fluor 488 conjugated goat anti mouse igg
    Treatment with iloprost prevents LPC-mediated vascular barrier disruption in the adult spinal cord. A , visualization of vascular leakage in the CNS. Upper panels, representative images of the thoracic spinal cord showing leakage of <t>Alexa</t> Fluor 555-conjugated cadaverine ( red ) into the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. The broken line represents the outline of the tissue. Scale bar, 1 mm. Middle panels, representative images of the thoracic spinal cord showing the distribution of Alexa Fluor 555-conjugated cadaverine ( red ) in the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. Sections were counterstained for CD31 (vascular endothelial cell marker, green ). Scale bar, 100 μm. Lower panels, representative images of cross-sections of the thoracic spinal cord showing leakage of endogenous <t>IgG</t> ( green ) into the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. Sections were vascular counterstained with DyLight 594-labeled L. esculentum lectin (vascular endothelial cell marker, red ). Scale bar, 100 μm. B , quantification of Evans Blue leakage in lesions of the spinal cord at 1 day after the operation. Values represent the mean ± S.E. of three independent experiments. **, p
    Alexa Fluor 488 Conjugated Goat Anti Mouse Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alexa fluor 488 conjugated goat anti mouse igg/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    96
    Thermo Fisher goat anti mouse igg conjugated to alexa fluor 488
    BrdU staining and its cellular localization in astrocyte cultures. Astrocyte cultures were incubated with solvent (A,D) , ADPβS (B,E) or 10% FBS, a positive control for cell proliferation (C,F) , for 48 h. BrdU (100 μM) was added to the medium for the last 24 h and then stained with rabbit anti-BrdU <t>(Alexa</t> Fluor 594, red) to visualize BrdU incorporation. Microglia were then co-labeled with mouse anti-Cd11b ( A–C ; Alexa Fluor 488, green) and astrocytes, with mouse anti-GFAP ( D–F ; Alexa Fluor 488, green). BrdU positive nuclei co-localize mainly with astrocytes, showing they are the main proliferating cells within the astrocyte cultures. Representative images from 3 different cultures. Scale bar: 100 μm.
    Goat Anti Mouse Igg Conjugated To Alexa Fluor 488, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse igg conjugated to alexa fluor 488/product/Thermo Fisher
    Average 96 stars, based on 1 article reviews
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    99
    Thermo Fisher alexa fluor 488 conjugated goat anti mouse igg secondary antibody
    Binding of DENV NS1 to Mouse Tissues Cryo-sections of mouse (A) lung, (B) liver, and (C) intestine were incubated with serum-free supernatants from BHK DENV-2 Rep or BHK cells for 1 h at room temperature. After extensive washing, bound NS1 was detected by a mixture of NS1 mAbs (1A4, 1F11, 2G6, 1B2) followed by Cy3-conjugated goat anti-mouse <t>IgG.</t> Co-staining with endothelial cell marker was subsequently performed by incubating the sections with rat anti-mouse CD31 (PECAM-1) followed by <t>Alexa</t> Fluor 488-conjugated goat anti-rat IgG. Nuclei were stained with a DNA-specific dye TO-PRO-3. Sections incubated with DENV NS1 followed by an isotype control Ab served as a negative control. Analysis was performed by confocal microscopy. White arrow and yellow arrowhead denote the layer of endothelial cells in the lumen and the outer layer of the adventitia of pulmonary vessel, respectively.
    Alexa Fluor 488 Conjugated Goat Anti Mouse Igg Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alexa fluor 488 conjugated goat anti mouse igg secondary antibody/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alexa fluor 488 conjugated goat anti mouse igg secondary antibody - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

    Image Search Results


    Chemical structures of the compounds tested and visualization of SARS-CoV-2-infected Vero cells. ( A ) Chemical structures of gemcitabine, 2FdC, and remdesivir. ( B ) MTT-based cytotoxicity assay of gemcitabine at different concentrations and time points. Vero cells were treated with increasing concentrations of gemcitabine for 24 h (black square) or 48 h (gray square). Percentage cell viability was measured by using MTT, in which mock-treated cells served as a control (100%). ( C ) Fluorescein diacetate-based cytotoxicity assay of gemcitabine. Vero cells were treated with increasing concentrations of gemcitabine for 24 (black square) and 48 h (gray square). Percentage cell viability was measured by addition of fluorescein diacetate, in which mock-treated cells served as a control (100%). ( D ) Fluorescein diacetate-based cytotoxicity assay of a delivery vehicle. Increasing concentrations of DMSO, 0.2, 0.6 and 1.8% ( v / v ), that were identically included in 100, 300 and 900 μM gemcitabine shown in ( C ), were treated to Vero cells for 24 (black bar) and 48 h (gray bar). Values in ( B – D ) are means ± standard deviations from thee three independent experiments. ( E ) Visualization of SARS-CoV-2 infection. Vero cells were mock-infected (Mock) or infected with SARS-CoV-2 at a multiplicity of infection (MOI) of 0.02 for 24 h. Viral spike (S) protein was probed with mouse anti-S antibody and Alexa Fluor 488-conjugated goat anti-mouse antibody (green). Cellular nuclei were counterstained with DAPI (blue). Magnification ×20.

    Journal: International Journal of Molecular Sciences

    Article Title: Comparison of Antiviral Activity of Gemcitabine with 2′-Fluoro-2′-Deoxycytidine and Combination Therapy with Remdesivir against SARS-CoV-2

    doi: 10.3390/ijms22041581

    Figure Lengend Snippet: Chemical structures of the compounds tested and visualization of SARS-CoV-2-infected Vero cells. ( A ) Chemical structures of gemcitabine, 2FdC, and remdesivir. ( B ) MTT-based cytotoxicity assay of gemcitabine at different concentrations and time points. Vero cells were treated with increasing concentrations of gemcitabine for 24 h (black square) or 48 h (gray square). Percentage cell viability was measured by using MTT, in which mock-treated cells served as a control (100%). ( C ) Fluorescein diacetate-based cytotoxicity assay of gemcitabine. Vero cells were treated with increasing concentrations of gemcitabine for 24 (black square) and 48 h (gray square). Percentage cell viability was measured by addition of fluorescein diacetate, in which mock-treated cells served as a control (100%). ( D ) Fluorescein diacetate-based cytotoxicity assay of a delivery vehicle. Increasing concentrations of DMSO, 0.2, 0.6 and 1.8% ( v / v ), that were identically included in 100, 300 and 900 μM gemcitabine shown in ( C ), were treated to Vero cells for 24 (black bar) and 48 h (gray bar). Values in ( B – D ) are means ± standard deviations from thee three independent experiments. ( E ) Visualization of SARS-CoV-2 infection. Vero cells were mock-infected (Mock) or infected with SARS-CoV-2 at a multiplicity of infection (MOI) of 0.02 for 24 h. Viral spike (S) protein was probed with mouse anti-S antibody and Alexa Fluor 488-conjugated goat anti-mouse antibody (green). Cellular nuclei were counterstained with DAPI (blue). Magnification ×20.

    Article Snippet: Viral S protein was probed using anti-S antibody (Cat. No., GTX632604; Genetex, Irvine, CA, USA) and Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Invitrogen, Carlsbad, CA, USA), while cellular nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; Invitrogen).

    Techniques: Infection, MTT Assay, Cytotoxicity Assay

    Treatment with iloprost prevents LPC-mediated vascular barrier disruption in the adult spinal cord. A , visualization of vascular leakage in the CNS. Upper panels, representative images of the thoracic spinal cord showing leakage of Alexa Fluor 555-conjugated cadaverine ( red ) into the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. The broken line represents the outline of the tissue. Scale bar, 1 mm. Middle panels, representative images of the thoracic spinal cord showing the distribution of Alexa Fluor 555-conjugated cadaverine ( red ) in the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. Sections were counterstained for CD31 (vascular endothelial cell marker, green ). Scale bar, 100 μm. Lower panels, representative images of cross-sections of the thoracic spinal cord showing leakage of endogenous IgG ( green ) into the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. Sections were vascular counterstained with DyLight 594-labeled L. esculentum lectin (vascular endothelial cell marker, red ). Scale bar, 100 μm. B , quantification of Evans Blue leakage in lesions of the spinal cord at 1 day after the operation. Values represent the mean ± S.E. of three independent experiments. **, p

    Journal: The Journal of Biological Chemistry

    Article Title: Prostacyclin Prevents Pericyte Loss and Demyelination Induced by Lysophosphatidylcholine in the Central Nervous System *

    doi: 10.1074/jbc.M114.587253

    Figure Lengend Snippet: Treatment with iloprost prevents LPC-mediated vascular barrier disruption in the adult spinal cord. A , visualization of vascular leakage in the CNS. Upper panels, representative images of the thoracic spinal cord showing leakage of Alexa Fluor 555-conjugated cadaverine ( red ) into the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. The broken line represents the outline of the tissue. Scale bar, 1 mm. Middle panels, representative images of the thoracic spinal cord showing the distribution of Alexa Fluor 555-conjugated cadaverine ( red ) in the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. Sections were counterstained for CD31 (vascular endothelial cell marker, green ). Scale bar, 100 μm. Lower panels, representative images of cross-sections of the thoracic spinal cord showing leakage of endogenous IgG ( green ) into the spinal cord 1 day after LPC injection. Iloprost treatment was initiated immediately after LPC injection. Sections were vascular counterstained with DyLight 594-labeled L. esculentum lectin (vascular endothelial cell marker, red ). Scale bar, 100 μm. B , quantification of Evans Blue leakage in lesions of the spinal cord at 1 day after the operation. Values represent the mean ± S.E. of three independent experiments. **, p

    Article Snippet: Sections were incubated with Alexa Fluor 488-conjugated goat anti-mouse IgG (1:500, Invitrogen, catalog no. A-11001) and DyLight 594-labeled L. esculentum (tomato) lectin (1:100) for 1 h at room temperature.

    Techniques: Injection, Marker, Labeling

    BrdU staining and its cellular localization in astrocyte cultures. Astrocyte cultures were incubated with solvent (A,D) , ADPβS (B,E) or 10% FBS, a positive control for cell proliferation (C,F) , for 48 h. BrdU (100 μM) was added to the medium for the last 24 h and then stained with rabbit anti-BrdU (Alexa Fluor 594, red) to visualize BrdU incorporation. Microglia were then co-labeled with mouse anti-Cd11b ( A–C ; Alexa Fluor 488, green) and astrocytes, with mouse anti-GFAP ( D–F ; Alexa Fluor 488, green). BrdU positive nuclei co-localize mainly with astrocytes, showing they are the main proliferating cells within the astrocyte cultures. Representative images from 3 different cultures. Scale bar: 100 μm.

    Journal: Frontiers in Pharmacology

    Article Title: Microglia P2Y13 Receptors Prevent Astrocyte Proliferation Mediated by P2Y1 Receptors

    doi: 10.3389/fphar.2018.00418

    Figure Lengend Snippet: BrdU staining and its cellular localization in astrocyte cultures. Astrocyte cultures were incubated with solvent (A,D) , ADPβS (B,E) or 10% FBS, a positive control for cell proliferation (C,F) , for 48 h. BrdU (100 μM) was added to the medium for the last 24 h and then stained with rabbit anti-BrdU (Alexa Fluor 594, red) to visualize BrdU incorporation. Microglia were then co-labeled with mouse anti-Cd11b ( A–C ; Alexa Fluor 488, green) and astrocytes, with mouse anti-GFAP ( D–F ; Alexa Fluor 488, green). BrdU positive nuclei co-localize mainly with astrocytes, showing they are the main proliferating cells within the astrocyte cultures. Representative images from 3 different cultures. Scale bar: 100 μm.

    Article Snippet: Drugs and Antibodies The following antibodies and drugs were used: goat anti-mouse IgG conjugated to Alexa Fluor 488 from Invitrogen (Barcelona, Spain); rabbit polyclonal anti-P2Y13 from Alomone Laboratories (Jerusalem, Israel); mouse monoclonal anti-CD11b, and goat anti-rabbit IgG conjugated to horseradish peroxidase from Santa Cruz Biotechnology (Santa Cruz, CA, United States); 5-bromo-20-deoxyuridine (BrdU), rabbit polyclonal anti-BrdU, rabbit polyclonal anti-α tubulin, goat anti-rabbit IgG conjugated to Alexa Fluor 594 from Abcam (Cambridge, United Kingdom); rabbit and mouse anti-glial fibrillary acidic protein (anti-GFAP), recombinant rat interleukin-1β, rabbit polyclonal anti-interleukin-1β antibody, adenosine 5’-O -(3-thio)-diphosphate tetralithium (ADPβS), 2′-(4-hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5′-bi-1H-benzimidazole trihydrochloride hydrate(Hoechst 33258), penicillin and streptomycin from Sigma-Aldrich (Sintra, Portugal); 2-(propylthio)adenosine-5′-O -(β,γ-difluoromethylene)triphosphate tetrasodium (AR-C66096), 2-[(2-chloro-5-nitrophenyl) azo]-5-hydroxy-6-methyl-3-[(phos-phonooxy)methyl]-4-pyridinecarboxaldehyde disodium (MRS 2211) and (1R∗ ,2S∗ )-4-[2-Iodo-6-(methylamino)-9H-purin-9-yl]-2-(phosphonooxy)bicyclo[3.1.0]hexane-1-methanol dihydro-gen phosphate ester tetraammonium (MRS 2500) from Tocris (Bristol, United Kingdom); methyl -[3 H]-thymidine (specific activity 80–86 Ci.mmol-1 ) and enhanced chemiluminescence (ECL) Western blotting system from Amersham Biosciences (Lisbon, Portugal); goat polyclonal IL-1α antibody, mouse polyclonal TNF-α antibody from ThermoFisher Scientific (Lisbon, Portugal).

    Techniques: BrdU Staining, Incubation, Positive Control, Staining, BrdU Incorporation Assay, Labeling

    Binding of DENV NS1 to Mouse Tissues Cryo-sections of mouse (A) lung, (B) liver, and (C) intestine were incubated with serum-free supernatants from BHK DENV-2 Rep or BHK cells for 1 h at room temperature. After extensive washing, bound NS1 was detected by a mixture of NS1 mAbs (1A4, 1F11, 2G6, 1B2) followed by Cy3-conjugated goat anti-mouse IgG. Co-staining with endothelial cell marker was subsequently performed by incubating the sections with rat anti-mouse CD31 (PECAM-1) followed by Alexa Fluor 488-conjugated goat anti-rat IgG. Nuclei were stained with a DNA-specific dye TO-PRO-3. Sections incubated with DENV NS1 followed by an isotype control Ab served as a negative control. Analysis was performed by confocal microscopy. White arrow and yellow arrowhead denote the layer of endothelial cells in the lumen and the outer layer of the adventitia of pulmonary vessel, respectively.

    Journal: PLoS Pathogens

    Article Title: Secreted NS1 of Dengue Virus Attaches to the Surface of Cells via Interactions with Heparan Sulfate and Chondroitin Sulfate E

    doi: 10.1371/journal.ppat.0030183

    Figure Lengend Snippet: Binding of DENV NS1 to Mouse Tissues Cryo-sections of mouse (A) lung, (B) liver, and (C) intestine were incubated with serum-free supernatants from BHK DENV-2 Rep or BHK cells for 1 h at room temperature. After extensive washing, bound NS1 was detected by a mixture of NS1 mAbs (1A4, 1F11, 2G6, 1B2) followed by Cy3-conjugated goat anti-mouse IgG. Co-staining with endothelial cell marker was subsequently performed by incubating the sections with rat anti-mouse CD31 (PECAM-1) followed by Alexa Fluor 488-conjugated goat anti-rat IgG. Nuclei were stained with a DNA-specific dye TO-PRO-3. Sections incubated with DENV NS1 followed by an isotype control Ab served as a negative control. Analysis was performed by confocal microscopy. White arrow and yellow arrowhead denote the layer of endothelial cells in the lumen and the outer layer of the adventitia of pulmonary vessel, respectively.

    Article Snippet: After three washes, sections were incubated with Alexa Fluor 488 conjugated goat anti-mouse IgG secondary antibody (Invitrogen) and followed by 1-h incubation at room temperature with rabbit anti-human CD31 (PECAM) (Santa Cruz Biotechnology) at the dilution of 1:10.

    Techniques: Binding Assay, Incubation, Staining, Marker, Negative Control, Confocal Microscopy

    Binding of DENV NS1 to Human Lung Cryo-sections of human lung tissue were incubated with 20 μg/ml of purified DENV-2 NS1 or BSA for 1 h at room temperature. Bound NS1 was detected by staining with a mixture of DENV NS1 mAbs (1A4, 1F11, 2G6, 1B2) followed by Alexa Fluor 488 conjugated with goat anti-mouse IgG. Subsequently, the sections were co-stained with rabbit anti-human CD31 (PECAM-1) followed by Cy3-conjugated donkey anti-rabbit IgG. Nuclei were stained with a DNA-specific dye (Hoechst) and the sections were analyzed by confocal microscopy. Sections incubated with purified DENV NS1 and stained with isotype mAbs served as a negative control.

    Journal: PLoS Pathogens

    Article Title: Secreted NS1 of Dengue Virus Attaches to the Surface of Cells via Interactions with Heparan Sulfate and Chondroitin Sulfate E

    doi: 10.1371/journal.ppat.0030183

    Figure Lengend Snippet: Binding of DENV NS1 to Human Lung Cryo-sections of human lung tissue were incubated with 20 μg/ml of purified DENV-2 NS1 or BSA for 1 h at room temperature. Bound NS1 was detected by staining with a mixture of DENV NS1 mAbs (1A4, 1F11, 2G6, 1B2) followed by Alexa Fluor 488 conjugated with goat anti-mouse IgG. Subsequently, the sections were co-stained with rabbit anti-human CD31 (PECAM-1) followed by Cy3-conjugated donkey anti-rabbit IgG. Nuclei were stained with a DNA-specific dye (Hoechst) and the sections were analyzed by confocal microscopy. Sections incubated with purified DENV NS1 and stained with isotype mAbs served as a negative control.

    Article Snippet: After three washes, sections were incubated with Alexa Fluor 488 conjugated goat anti-mouse IgG secondary antibody (Invitrogen) and followed by 1-h incubation at room temperature with rabbit anti-human CD31 (PECAM) (Santa Cruz Biotechnology) at the dilution of 1:10.

    Techniques: Binding Assay, Incubation, Purification, Staining, Confocal Microscopy, Negative Control