alexa fluor 488 conjugated goat anti mouse igg secondary 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 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.
    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 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, 2021-06
    99/100 stars

    Images

    1) Product Images from "Secreted NS1 of Dengue Virus Attaches to the Surface of Cells via Interactions with Heparan Sulfate and Chondroitin Sulfate E"

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

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.0030183

    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.
    Figure Legend 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.

    Techniques Used: 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.
    Figure Legend 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.

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

    2) Product Images from "Plant-Produced Antigen Displaying Virus-Like Particles Evokes Potent Antibody Responses against West Nile Virus in Mice"

    Article Title: Plant-Produced Antigen Displaying Virus-Like Particles Evokes Potent Antibody Responses against West Nile Virus in Mice

    Journal: Vaccines

    doi: 10.3390/vaccines9010060

    Competitive binding between the E16 mAb and antibodies in anti-HBcAg-wDIII serum to wDIII displayed on the yeast cell surface. wDIII-displaying yeast cells were pre-incubated with PBS ( A ) or week-11 pooled sera (1:1000 dilution) from mice either injected with PBS ( B ) or HBcAg-wDIII VLP ( C ). The E16 mAb was then incubated with yeast cells. The specific binding between E16 and yeast-displayed wDIII was measured by staining with an Alexa Fluor 488-conjugated goat anti-human secondary antibody and processing by flow cytometry.
    Figure Legend Snippet: Competitive binding between the E16 mAb and antibodies in anti-HBcAg-wDIII serum to wDIII displayed on the yeast cell surface. wDIII-displaying yeast cells were pre-incubated with PBS ( A ) or week-11 pooled sera (1:1000 dilution) from mice either injected with PBS ( B ) or HBcAg-wDIII VLP ( C ). The E16 mAb was then incubated with yeast cells. The specific binding between E16 and yeast-displayed wDIII was measured by staining with an Alexa Fluor 488-conjugated goat anti-human secondary antibody and processing by flow cytometry.

    Techniques Used: Binding Assay, Incubation, Mouse Assay, Injection, Staining, Flow Cytometry

    3) Product Images from "Identification of a Novel Nucleocytoplasmic Shuttling RNA Helicase of Trypanosomes"

    Article Title: Identification of a Novel Nucleocytoplasmic Shuttling RNA Helicase of Trypanosomes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0109521

    Localization of Hel45 after actinomycin D treatment in T. cruzi . Detection of exogenous Hel45 (A) tagged with PTP at the NT by indirect immunofluorescence with an anti-ProtA antibody and of mRNA (B) by fluorescence in situ hybridization (FISH) with a digoxigenin-conjugated oligo(dT) probe in T. cruzi after treatment with 50 µg/ml actinomycin D (ACTD) for 24 hours. Probe detection was carried out by indirect immunofluorescence with anti-DIG mouse monoclonal antibodies (Sigma-Aldrich, 1∶300 dilution) followed by secondary Alexa Fluor 488-conjugated antibodies (1∶600 dilution). As a control, 100 µg/ml RNase A was incubated with the parasites before probe hybridization (RNase A). DAPI = DNA stained with DAPI. Hel45 = localization of tagged Hel45. MERGE = merged images for DAPI staining and Hel45 or mRNA localization. N = nucleus. K = kinetoplast. Arrows = parasites with nuclear accumulation of tagged Hel45. Bar = 5 µm.
    Figure Legend Snippet: Localization of Hel45 after actinomycin D treatment in T. cruzi . Detection of exogenous Hel45 (A) tagged with PTP at the NT by indirect immunofluorescence with an anti-ProtA antibody and of mRNA (B) by fluorescence in situ hybridization (FISH) with a digoxigenin-conjugated oligo(dT) probe in T. cruzi after treatment with 50 µg/ml actinomycin D (ACTD) for 24 hours. Probe detection was carried out by indirect immunofluorescence with anti-DIG mouse monoclonal antibodies (Sigma-Aldrich, 1∶300 dilution) followed by secondary Alexa Fluor 488-conjugated antibodies (1∶600 dilution). As a control, 100 µg/ml RNase A was incubated with the parasites before probe hybridization (RNase A). DAPI = DNA stained with DAPI. Hel45 = localization of tagged Hel45. MERGE = merged images for DAPI staining and Hel45 or mRNA localization. N = nucleus. K = kinetoplast. Arrows = parasites with nuclear accumulation of tagged Hel45. Bar = 5 µm.

    Techniques Used: Immunofluorescence, Fluorescence, In Situ Hybridization, Fluorescence In Situ Hybridization, Incubation, Hybridization, Staining

    Related Articles

    Staining:

    Article Title: An immunotoxin targeting Ebola virus glycoprotein inhibits Ebola virus production from infected cells
    Article Snippet: At various times post-inoculation, cell plates were fixed with 10% neutral buffered formalin (NBF, Thermo Fisher Scientific) for 24 h and then transferred from the BSL-4 to a BSL-2 laboratory. .. Without the permeabilization step, the plates were stained with mouse anti-EBOV GP1,2 mAb 6D8, followed by secondary Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Thermo Fisher Scientific). .. Cell nuclei were stained with Hoechst 33342 dye (Thermo Fisher Scientific).

    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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  • 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, 2021-06
    99/100 stars
      Buy from Supplier

    86
    Thermo Fisher secondary antibody goat anti mouse igg alexa fluor 488 conjugate
    Pneumococcal lipoproteins are highly abundant on the pneumococcal surface. (A) In immunoblots, the specificity of antisera derived from intraperitoneal immunisations of CD-1 mice ( n = 6) with recombinant lipoproteins was assessed. Therefore, the wild-type strain S. pneumoniae D39Δ cps and the corresponding isogenic lipoprotein deficient mutants (2 × 10 8 bacteria per lane) were used. Enolase was detected with a rabbit anti-enolase serum and served as a loading control. (B) <t>IgG</t> antibody titrations were performed by incubating equimolar amounts of recombinant proteins with serial dilutions of isolated polyclonal IgGs. Detection was carried out using a peroxidase-coupled goat anti-mouse IgG followed by incubation with OPD as a substrate and absorbance was measured at 492 nm. Titrations were performed at least three times and the error bars represent the SEM. (C,D) Using the equation for the hyperbolic regression curve ( y [ A b s ] = B m a x · x K d + x , Bmax, maximal binding; Kd, concentration for half maximal binding) an initial IgG concentration was calculated in the linear dynamic range. The polyclonal IgGs with equal contents of IgG specific for each lipoprotein were therefore applied to enable the comparison of their surface abundances. In a flow cytometric approach, D39Δ cps (C) and D39 (D) were incubated with the appropriate calculated concentration of IgG and concentrations 5-, 10-, 20-, and 50-fold greater to analyse the surface abundance of the selected lipoproteins. Antibody binding was detected using a goat anti-mouse <t>Alexa</t> Fluor® 488-coupled secondary antibody. The percentage of positive gated events is depicted in the graphs, thereby indicating the proportion of wild-type bacteria positive for the binding of the respective anti-lipoprotein IgGs. The mean values of at least three independent experiments are shown, with error bars corresponding to SEM.
    Secondary Antibody Goat Anti Mouse Igg Alexa Fluor 488 Conjugate, 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/secondary antibody goat anti mouse igg alexa fluor 488 conjugate/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    secondary antibody goat anti mouse igg alexa fluor 488 conjugate - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    Image Search Results


    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

    6D8-PE38 recombinant immunotoxin specially binds to EBOV GP 1,2 -expressing cells. 6D8-PE38 RIT was labeled with Alexa Fluor 488. Vero E6 cells were transfected with pCAGGS-EBOV-GP 1,2 or control pCAGGS-MARV-GP 1,2 plasmids. After 48 h, transfected cells were incubated with the indicated concentrations of labeled 6D8-PE38 RIT. Cell-bound RIT was detected by flow cytometry. FSC-H: forward scatter height.

    Journal: PLoS ONE

    Article Title: An immunotoxin targeting Ebola virus glycoprotein inhibits Ebola virus production from infected cells

    doi: 10.1371/journal.pone.0245024

    Figure Lengend Snippet: 6D8-PE38 recombinant immunotoxin specially binds to EBOV GP 1,2 -expressing cells. 6D8-PE38 RIT was labeled with Alexa Fluor 488. Vero E6 cells were transfected with pCAGGS-EBOV-GP 1,2 or control pCAGGS-MARV-GP 1,2 plasmids. After 48 h, transfected cells were incubated with the indicated concentrations of labeled 6D8-PE38 RIT. Cell-bound RIT was detected by flow cytometry. FSC-H: forward scatter height.

    Article Snippet: Without the permeabilization step, the plates were stained with mouse anti-EBOV GP1,2 mAb 6D8, followed by secondary Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Thermo Fisher Scientific).

    Techniques: Recombinant, Expressing, Labeling, Transfection, Incubation, Flow Cytometry

    Recognition of EBOV GP 1,2 expressed on the cell surface by mAb 6D8. Vero E6 cells were transfected with plasmids pCAGGS-EBOV-GP 1, 2 , pCAGGS-MARV-GP 1,2 , or pCAGGS. After 48 h, cells were dissociated with enzyme-free cell-dissociation buffer and stained with mAb 6D8 or control mouse immunoglobulin G (IgG), followed by Alexa Fluor 488-conjugated goat anti-mouse IgG antibody. FSC-A: forward scatter area.

    Journal: PLoS ONE

    Article Title: An immunotoxin targeting Ebola virus glycoprotein inhibits Ebola virus production from infected cells

    doi: 10.1371/journal.pone.0245024

    Figure Lengend Snippet: Recognition of EBOV GP 1,2 expressed on the cell surface by mAb 6D8. Vero E6 cells were transfected with plasmids pCAGGS-EBOV-GP 1, 2 , pCAGGS-MARV-GP 1,2 , or pCAGGS. After 48 h, cells were dissociated with enzyme-free cell-dissociation buffer and stained with mAb 6D8 or control mouse immunoglobulin G (IgG), followed by Alexa Fluor 488-conjugated goat anti-mouse IgG antibody. FSC-A: forward scatter area.

    Article Snippet: Without the permeabilization step, the plates were stained with mouse anti-EBOV GP1,2 mAb 6D8, followed by secondary Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Thermo Fisher Scientific).

    Techniques: Transfection, Staining

    Pneumococcal lipoproteins are highly abundant on the pneumococcal surface. (A) In immunoblots, the specificity of antisera derived from intraperitoneal immunisations of CD-1 mice ( n = 6) with recombinant lipoproteins was assessed. Therefore, the wild-type strain S. pneumoniae D39Δ cps and the corresponding isogenic lipoprotein deficient mutants (2 × 10 8 bacteria per lane) were used. Enolase was detected with a rabbit anti-enolase serum and served as a loading control. (B) IgG antibody titrations were performed by incubating equimolar amounts of recombinant proteins with serial dilutions of isolated polyclonal IgGs. Detection was carried out using a peroxidase-coupled goat anti-mouse IgG followed by incubation with OPD as a substrate and absorbance was measured at 492 nm. Titrations were performed at least three times and the error bars represent the SEM. (C,D) Using the equation for the hyperbolic regression curve ( y [ A b s ] = B m a x · x K d + x , Bmax, maximal binding; Kd, concentration for half maximal binding) an initial IgG concentration was calculated in the linear dynamic range. The polyclonal IgGs with equal contents of IgG specific for each lipoprotein were therefore applied to enable the comparison of their surface abundances. In a flow cytometric approach, D39Δ cps (C) and D39 (D) were incubated with the appropriate calculated concentration of IgG and concentrations 5-, 10-, 20-, and 50-fold greater to analyse the surface abundance of the selected lipoproteins. Antibody binding was detected using a goat anti-mouse Alexa Fluor® 488-coupled secondary antibody. The percentage of positive gated events is depicted in the graphs, thereby indicating the proportion of wild-type bacteria positive for the binding of the respective anti-lipoprotein IgGs. The mean values of at least three independent experiments are shown, with error bars corresponding to SEM.

    Journal: Frontiers in Immunology

    Article Title: Intranasal Vaccination With Lipoproteins Confers Protection Against Pneumococcal Colonisation

    doi: 10.3389/fimmu.2018.02405

    Figure Lengend Snippet: Pneumococcal lipoproteins are highly abundant on the pneumococcal surface. (A) In immunoblots, the specificity of antisera derived from intraperitoneal immunisations of CD-1 mice ( n = 6) with recombinant lipoproteins was assessed. Therefore, the wild-type strain S. pneumoniae D39Δ cps and the corresponding isogenic lipoprotein deficient mutants (2 × 10 8 bacteria per lane) were used. Enolase was detected with a rabbit anti-enolase serum and served as a loading control. (B) IgG antibody titrations were performed by incubating equimolar amounts of recombinant proteins with serial dilutions of isolated polyclonal IgGs. Detection was carried out using a peroxidase-coupled goat anti-mouse IgG followed by incubation with OPD as a substrate and absorbance was measured at 492 nm. Titrations were performed at least three times and the error bars represent the SEM. (C,D) Using the equation for the hyperbolic regression curve ( y [ A b s ] = B m a x · x K d + x , Bmax, maximal binding; Kd, concentration for half maximal binding) an initial IgG concentration was calculated in the linear dynamic range. The polyclonal IgGs with equal contents of IgG specific for each lipoprotein were therefore applied to enable the comparison of their surface abundances. In a flow cytometric approach, D39Δ cps (C) and D39 (D) were incubated with the appropriate calculated concentration of IgG and concentrations 5-, 10-, 20-, and 50-fold greater to analyse the surface abundance of the selected lipoproteins. Antibody binding was detected using a goat anti-mouse Alexa Fluor® 488-coupled secondary antibody. The percentage of positive gated events is depicted in the graphs, thereby indicating the proportion of wild-type bacteria positive for the binding of the respective anti-lipoprotein IgGs. The mean values of at least three independent experiments are shown, with error bars corresponding to SEM.

    Article Snippet: The bacteria were washed with PBS and stained using secondary antibody goat anti-mouse IgG Alexa-Fluor-488 conjugate (1:1,000; Thermo Fisher Scientific).

    Techniques: Western Blot, Derivative Assay, Mouse Assay, Recombinant, Isolation, Incubation, Binding Assay, Concentration Assay, Flow Cytometry