goat anti-mouse Search Results


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  • 99
    LI-COR irdye 800cw goat anti mouse igg h l
    Irdye 800cw Goat Anti Mouse Igg H L, supplied by LI-COR, used in various techniques. Bioz Stars score: 99/100, based on 2660 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse igg h l cross adsorbed secondary antibody
    Goat Anti Mouse Igg H L Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14216 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse igg h l highly cross adsorbed secondary antibody
    Goat Anti Mouse Igg H L Highly Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 10039 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse igg
    Goat Anti Mouse Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9641 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse
    Goat Anti Mouse, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 9544 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher secondary antibodies
    Secondary Antibodies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 16006 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology goat anti mouse igg hrp
    Goat Anti Mouse Igg Hrp, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 4245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Jackson Immuno peroxidase affinipure goat anti mouse igg
    Peroxidase Affinipure Goat Anti Mouse Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 99/100, based on 3302 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore goat anti mouse igg
    Goat Anti Mouse Igg, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3554 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Jackson Immuno goat anti mouse igg
    Goat Anti Mouse Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 99/100, based on 3801 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology goat anti mouse
    Goat Anti Mouse, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 97/100, based on 3727 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti mouse igg whole molecule peroxidase antibody
    Anti Mouse Igg Whole Molecule Peroxidase Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5324 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse igg1 cross adsorbed secondary antibody
    Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP <t>IgG1</t> in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test
    Goat Anti Mouse Igg1 Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1214 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SouthernBiotech goat anti mouse igg1
    Induction of anti-HA humoral and cellular responses in mice vaccinated with VLP-HA/VSP-G. a – e Mice were orally vaccinated with VLPs and the humoral response against HA in different biological samples was evaluated. Levels of total serum anti-HA IgG ( a ), <t>IgG1</t> and IgG2a ( b ), n = 8 from two independent experiments. Levels of IgA anti-HA in fecal extracts and BAL, n = 6 from two independent experiments ( c ). Levels of serum IgG anti-HA measured at different times post immunization, n = 6 from two independent experiments ( d ). Levels of serum IgG anti-HA in animals immunized with a mixture of VLP-HA plus VLP-VSP-G, n = 6 from two independent experiments ( e ). f Levels of serum IgG and fecal IgA in mice vaccinated with VLPs via oral or subcutaneous (s.c.) route, n = 8 from two independent experiments. g Mice were orally immunized with four weekly doses of 50 μg of recombinant ΔVSP1267 or vehicle. One week after the last immunization the presence of anti-VSP1267 antibodies in blood and fecal samples was checked, then these animals were orally immunized with VLP-HA/VSP-G, according to the protocol previously used. One week after the last VLP dose, the levels of serum IgG and fecal IgA anti-HA Abs were measured, n = 6 from two independent experiments. h , i Mice were orally vaccinated with VLPs and the cellular response against HA was evaluated. The frequency of HA-specific IFN-γ-secreting T cells (spot-forming colonies, SFC) was determined after antigen-specific re-stimulation of splenocytes, n = 7 from two independent experiments ( h ). Cytokines were measured in splenocyte supernatants and only those cytokines with detectable levels are shown, n = 6 from two independent experiments ( i ). Data were analyzed by one-way ANOVA and Tukey’s multiple comparison test ( a – f , h , i ) or by two-way ANOVA and Bonferroni post-tests ( g ). Values represent mean ± s.e.m. * p
    Goat Anti Mouse Igg1, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 99/100, based on 707 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam goat anti mouse igg h l hrp
    Induction of anti-HA humoral and cellular responses in mice vaccinated with VLP-HA/VSP-G. a – e Mice were orally vaccinated with VLPs and the humoral response against HA in different biological samples was evaluated. Levels of total serum anti-HA IgG ( a ), <t>IgG1</t> and IgG2a ( b ), n = 8 from two independent experiments. Levels of IgA anti-HA in fecal extracts and BAL, n = 6 from two independent experiments ( c ). Levels of serum IgG anti-HA measured at different times post immunization, n = 6 from two independent experiments ( d ). Levels of serum IgG anti-HA in animals immunized with a mixture of VLP-HA plus VLP-VSP-G, n = 6 from two independent experiments ( e ). f Levels of serum IgG and fecal IgA in mice vaccinated with VLPs via oral or subcutaneous (s.c.) route, n = 8 from two independent experiments. g Mice were orally immunized with four weekly doses of 50 μg of recombinant ΔVSP1267 or vehicle. One week after the last immunization the presence of anti-VSP1267 antibodies in blood and fecal samples was checked, then these animals were orally immunized with VLP-HA/VSP-G, according to the protocol previously used. One week after the last VLP dose, the levels of serum IgG and fecal IgA anti-HA Abs were measured, n = 6 from two independent experiments. h , i Mice were orally vaccinated with VLPs and the cellular response against HA was evaluated. The frequency of HA-specific IFN-γ-secreting T cells (spot-forming colonies, SFC) was determined after antigen-specific re-stimulation of splenocytes, n = 7 from two independent experiments ( h ). Cytokines were measured in splenocyte supernatants and only those cytokines with detectable levels are shown, n = 6 from two independent experiments ( i ). Data were analyzed by one-way ANOVA and Tukey’s multiple comparison test ( a – f , h , i ) or by two-way ANOVA and Bonferroni post-tests ( g ). Values represent mean ± s.e.m. * p
    Goat Anti Mouse Igg H L Hrp, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 729 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse secondary antibody
    Induction of anti-HA humoral and cellular responses in mice vaccinated with VLP-HA/VSP-G. a – e Mice were orally vaccinated with VLPs and the humoral response against HA in different biological samples was evaluated. Levels of total serum anti-HA IgG ( a ), <t>IgG1</t> and IgG2a ( b ), n = 8 from two independent experiments. Levels of IgA anti-HA in fecal extracts and BAL, n = 6 from two independent experiments ( c ). Levels of serum IgG anti-HA measured at different times post immunization, n = 6 from two independent experiments ( d ). Levels of serum IgG anti-HA in animals immunized with a mixture of VLP-HA plus VLP-VSP-G, n = 6 from two independent experiments ( e ). f Levels of serum IgG and fecal IgA in mice vaccinated with VLPs via oral or subcutaneous (s.c.) route, n = 8 from two independent experiments. g Mice were orally immunized with four weekly doses of 50 μg of recombinant ΔVSP1267 or vehicle. One week after the last immunization the presence of anti-VSP1267 antibodies in blood and fecal samples was checked, then these animals were orally immunized with VLP-HA/VSP-G, according to the protocol previously used. One week after the last VLP dose, the levels of serum IgG and fecal IgA anti-HA Abs were measured, n = 6 from two independent experiments. h , i Mice were orally vaccinated with VLPs and the cellular response against HA was evaluated. The frequency of HA-specific IFN-γ-secreting T cells (spot-forming colonies, SFC) was determined after antigen-specific re-stimulation of splenocytes, n = 7 from two independent experiments ( h ). Cytokines were measured in splenocyte supernatants and only those cytokines with detectable levels are shown, n = 6 from two independent experiments ( i ). Data were analyzed by one-way ANOVA and Tukey’s multiple comparison test ( a – f , h , i ) or by two-way ANOVA and Bonferroni post-tests ( g ). Values represent mean ± s.e.m. * p
    Goat Anti Mouse Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1644 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 594 goat anti mouse igg
    CAT1/SLC7A1 protein acted as a cellular receptor and was required for BLV infection. A ) Confirmation of the absence of endogenous CAT1 expression in CHO-K1 cells. CHO-K1 cells were transfected with and without bCAT1/pEGFP-N1 or with pEGFP-N1 as a negative control. The cell lysates were prepared 48 h post-transfection and then subjected to Western blot analysis using anti-CAT1 (upper panel), anti-EGFP (middle panel), and anti–β-actin (lower panel) antibodies. CC81 cell lysates were used as a positive control. The positions and MW of CAT1-EGFP, CAT1, EGFP, and actin are indicated. B ) bCAT1/SLC7A1 expression associated with BLV cell-free and cell-to-cell infection. CHO-K1 cells were transfected with bCAT1/pEGFP-N1 expression plasmid with or without the BLV infectious molecular clone pBLV-IF2 or cocultured with supernatants from FLK-BLV cells or FLK-BLV cells at 4 h post-transfection. CHO-K1 cells were transfected with bCAT1/pEGFP-N1 or with pEGFP-N1 as a negative control. After 48 h incubation, the cells were fixed with 3.7% formaldehyde and stained with 10 µg/ml Hoechst 33342. EGFP-expressing syncytia were evaluated and quantitated using EVOS2 fluorescence microscopy. C ) Detection of CAT1 and viral protein Env in syncytium. The transfected CHO-K1 cells were labeled with an anti-gp51 mAb (BLV-1) followed by incubation with <t>Alexa</t> <t>Fluor</t> 594 goat anti-mouse <t>IgG</t> (red). Nuclei were stained with Hoechst 33342 (blue). CAT1-EGFP was visualized to determine CAT1 expression (green) in the cells.
    Alexa Fluor 594 Goat Anti Mouse Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2483 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Category Antibodies Secondary Antibody Goat Anti Mouse IgM H L peroxidase HRP conjugated Size 60μL Price 20 Synonyms Goat Anti Mouse IgM Reactivity Mouse Source Goat Immunogen Mouse IgM Host
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    N/A
    Category Antibodies Secondary Antibody Goat Anti Mouse IgG H L peroxidase HRP conjugated Size 60μL Price 20 Synonyms Goat Anti Mouse IgG Reactivity Mouse Source Goat Immunogen Mouse IgG Host
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    Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP IgG1 in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test

    Journal: Nature Communications

    Article Title: PRMT5 is essential for B cell development and germinal center dynamics

    doi: 10.1038/s41467-018-07884-6

    Figure Lengend Snippet: Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP IgG1 in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test

    Article Snippet: Plates with cells were incubated in a humid chamber 12 h at 37 °C, 5% CO2 , then washed 6× with PBS 0.01% Tween-20, followed by incubation with goat anti-mouse IgG1-HRP (A10551, Life Technologies, 1/2000) diluted in culture media for 2 h at RT.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Flow Cytometry, Cytometry, Infection, Staining, Two Tailed Test

    Induction of anti-HA humoral and cellular responses in mice vaccinated with VLP-HA/VSP-G. a – e Mice were orally vaccinated with VLPs and the humoral response against HA in different biological samples was evaluated. Levels of total serum anti-HA IgG ( a ), IgG1 and IgG2a ( b ), n = 8 from two independent experiments. Levels of IgA anti-HA in fecal extracts and BAL, n = 6 from two independent experiments ( c ). Levels of serum IgG anti-HA measured at different times post immunization, n = 6 from two independent experiments ( d ). Levels of serum IgG anti-HA in animals immunized with a mixture of VLP-HA plus VLP-VSP-G, n = 6 from two independent experiments ( e ). f Levels of serum IgG and fecal IgA in mice vaccinated with VLPs via oral or subcutaneous (s.c.) route, n = 8 from two independent experiments. g Mice were orally immunized with four weekly doses of 50 μg of recombinant ΔVSP1267 or vehicle. One week after the last immunization the presence of anti-VSP1267 antibodies in blood and fecal samples was checked, then these animals were orally immunized with VLP-HA/VSP-G, according to the protocol previously used. One week after the last VLP dose, the levels of serum IgG and fecal IgA anti-HA Abs were measured, n = 6 from two independent experiments. h , i Mice were orally vaccinated with VLPs and the cellular response against HA was evaluated. The frequency of HA-specific IFN-γ-secreting T cells (spot-forming colonies, SFC) was determined after antigen-specific re-stimulation of splenocytes, n = 7 from two independent experiments ( h ). Cytokines were measured in splenocyte supernatants and only those cytokines with detectable levels are shown, n = 6 from two independent experiments ( i ). Data were analyzed by one-way ANOVA and Tukey’s multiple comparison test ( a – f , h , i ) or by two-way ANOVA and Bonferroni post-tests ( g ). Values represent mean ± s.e.m. * p

    Journal: Nature Communications

    Article Title: Efficient oral vaccination by bioengineering virus-like particles with protozoan surface proteins

    doi: 10.1038/s41467-018-08265-9

    Figure Lengend Snippet: Induction of anti-HA humoral and cellular responses in mice vaccinated with VLP-HA/VSP-G. a – e Mice were orally vaccinated with VLPs and the humoral response against HA in different biological samples was evaluated. Levels of total serum anti-HA IgG ( a ), IgG1 and IgG2a ( b ), n = 8 from two independent experiments. Levels of IgA anti-HA in fecal extracts and BAL, n = 6 from two independent experiments ( c ). Levels of serum IgG anti-HA measured at different times post immunization, n = 6 from two independent experiments ( d ). Levels of serum IgG anti-HA in animals immunized with a mixture of VLP-HA plus VLP-VSP-G, n = 6 from two independent experiments ( e ). f Levels of serum IgG and fecal IgA in mice vaccinated with VLPs via oral or subcutaneous (s.c.) route, n = 8 from two independent experiments. g Mice were orally immunized with four weekly doses of 50 μg of recombinant ΔVSP1267 or vehicle. One week after the last immunization the presence of anti-VSP1267 antibodies in blood and fecal samples was checked, then these animals were orally immunized with VLP-HA/VSP-G, according to the protocol previously used. One week after the last VLP dose, the levels of serum IgG and fecal IgA anti-HA Abs were measured, n = 6 from two independent experiments. h , i Mice were orally vaccinated with VLPs and the cellular response against HA was evaluated. The frequency of HA-specific IFN-γ-secreting T cells (spot-forming colonies, SFC) was determined after antigen-specific re-stimulation of splenocytes, n = 7 from two independent experiments ( h ). Cytokines were measured in splenocyte supernatants and only those cytokines with detectable levels are shown, n = 6 from two independent experiments ( i ). Data were analyzed by one-way ANOVA and Tukey’s multiple comparison test ( a – f , h , i ) or by two-way ANOVA and Bonferroni post-tests ( g ). Values represent mean ± s.e.m. * p

    Article Snippet: The following horseradish peroxidase-conjugated antibodies were used: goat anti-mouse IgG1 (SouthernBiotech, Cat. # 1070-05, dilution 1/2000); goat anti-mouse IgG2 (SouthernBiotech, Cat. # 1080-05, dilution 1/2000); goat anti-mouse IgG (Molecular Probes, Cat. # G-21040, dilution 1/2000).

    Techniques: Mouse Assay, Recombinant

    CAT1/SLC7A1 protein acted as a cellular receptor and was required for BLV infection. A ) Confirmation of the absence of endogenous CAT1 expression in CHO-K1 cells. CHO-K1 cells were transfected with and without bCAT1/pEGFP-N1 or with pEGFP-N1 as a negative control. The cell lysates were prepared 48 h post-transfection and then subjected to Western blot analysis using anti-CAT1 (upper panel), anti-EGFP (middle panel), and anti–β-actin (lower panel) antibodies. CC81 cell lysates were used as a positive control. The positions and MW of CAT1-EGFP, CAT1, EGFP, and actin are indicated. B ) bCAT1/SLC7A1 expression associated with BLV cell-free and cell-to-cell infection. CHO-K1 cells were transfected with bCAT1/pEGFP-N1 expression plasmid with or without the BLV infectious molecular clone pBLV-IF2 or cocultured with supernatants from FLK-BLV cells or FLK-BLV cells at 4 h post-transfection. CHO-K1 cells were transfected with bCAT1/pEGFP-N1 or with pEGFP-N1 as a negative control. After 48 h incubation, the cells were fixed with 3.7% formaldehyde and stained with 10 µg/ml Hoechst 33342. EGFP-expressing syncytia were evaluated and quantitated using EVOS2 fluorescence microscopy. C ) Detection of CAT1 and viral protein Env in syncytium. The transfected CHO-K1 cells were labeled with an anti-gp51 mAb (BLV-1) followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red). Nuclei were stained with Hoechst 33342 (blue). CAT1-EGFP was visualized to determine CAT1 expression (green) in the cells.

    Journal: The FASEB Journal

    Article Title: CAT1/SLC7A1 acts as a cellular receptor for bovine leukemia virus infection

    doi: 10.1096/fj.201901528R

    Figure Lengend Snippet: CAT1/SLC7A1 protein acted as a cellular receptor and was required for BLV infection. A ) Confirmation of the absence of endogenous CAT1 expression in CHO-K1 cells. CHO-K1 cells were transfected with and without bCAT1/pEGFP-N1 or with pEGFP-N1 as a negative control. The cell lysates were prepared 48 h post-transfection and then subjected to Western blot analysis using anti-CAT1 (upper panel), anti-EGFP (middle panel), and anti–β-actin (lower panel) antibodies. CC81 cell lysates were used as a positive control. The positions and MW of CAT1-EGFP, CAT1, EGFP, and actin are indicated. B ) bCAT1/SLC7A1 expression associated with BLV cell-free and cell-to-cell infection. CHO-K1 cells were transfected with bCAT1/pEGFP-N1 expression plasmid with or without the BLV infectious molecular clone pBLV-IF2 or cocultured with supernatants from FLK-BLV cells or FLK-BLV cells at 4 h post-transfection. CHO-K1 cells were transfected with bCAT1/pEGFP-N1 or with pEGFP-N1 as a negative control. After 48 h incubation, the cells were fixed with 3.7% formaldehyde and stained with 10 µg/ml Hoechst 33342. EGFP-expressing syncytia were evaluated and quantitated using EVOS2 fluorescence microscopy. C ) Detection of CAT1 and viral protein Env in syncytium. The transfected CHO-K1 cells were labeled with an anti-gp51 mAb (BLV-1) followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red). Nuclei were stained with Hoechst 33342 (blue). CAT1-EGFP was visualized to determine CAT1 expression (green) in the cells.

    Article Snippet: Cells were labeled with an anti–BLV Env antibody followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red fluorescence) to detect cells expressing Env and staining with Hoechst 33342 (blue fluorescence) for detection of the nucleus.

    Techniques: Infection, Expressing, Transfection, Negative Control, Western Blot, Positive Control, Plasmid Preparation, Incubation, Staining, Fluorescence, Microscopy, Labeling

    Colocalization of CAT1/SLC7A1 and BLV Env protein in endomembrane compartments and at the cell membrane. bCAT1/SLC7A1 expression plasmid bCAT1/pEGFP-N1 was transfected into both FLK-BLV cells ( A ) and PK15/pBLV-IF2, which was stably transfected pBLV-IF2 ( B ); the transfected cells were grown on cover glasses and labeled with an anti-gp51 mAb (BLV-1) followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red). Nuclei were stained with Hoechst 33342 (blue). CAT1-EGFP was visualized to determine CAT1 expression (green) in the cells. The cells were evaluated, and the mean fluorescence intensities were visualized and analyzed along the line using an Olympus FV1000 laser-scanning confocal fluorescence microscope. White arrows and numbers indicate that positions of endomembrane compartments and the cell membrane.

    Journal: The FASEB Journal

    Article Title: CAT1/SLC7A1 acts as a cellular receptor for bovine leukemia virus infection

    doi: 10.1096/fj.201901528R

    Figure Lengend Snippet: Colocalization of CAT1/SLC7A1 and BLV Env protein in endomembrane compartments and at the cell membrane. bCAT1/SLC7A1 expression plasmid bCAT1/pEGFP-N1 was transfected into both FLK-BLV cells ( A ) and PK15/pBLV-IF2, which was stably transfected pBLV-IF2 ( B ); the transfected cells were grown on cover glasses and labeled with an anti-gp51 mAb (BLV-1) followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red). Nuclei were stained with Hoechst 33342 (blue). CAT1-EGFP was visualized to determine CAT1 expression (green) in the cells. The cells were evaluated, and the mean fluorescence intensities were visualized and analyzed along the line using an Olympus FV1000 laser-scanning confocal fluorescence microscope. White arrows and numbers indicate that positions of endomembrane compartments and the cell membrane.

    Article Snippet: Cells were labeled with an anti–BLV Env antibody followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red fluorescence) to detect cells expressing Env and staining with Hoechst 33342 (blue fluorescence) for detection of the nucleus.

    Techniques: Expressing, Plasmid Preparation, Transfection, Stable Transfection, Labeling, Incubation, Staining, Fluorescence, Microscopy

    CAT1/SLC7A1 did not have species specificity with all demonstrating susceptibility to BLV infection. A ) CAT1/SLC7A1s from 9 different animal species did not have species specificity for BLV infection. The pEGFP-N1–expression plasmids encoding different CAT1/SLC7A1 proteins were derived using various animal species, including cattle (bovine), sheep (ovine), human, monkey (primate), pig (porcine), cat (feline), rat, mouse, and Chinese hamster. The plasmids were constructed and transfected into CHO-K1 cells. After 4 h transfection, the cells were added to culture supernatants of FLK-BLV cells and cultured for 48 h. The cells were then fixed with 3.7% formaldehyde and stained with 10 µg/ml Hoechst 33342. EGFP-expressing syncytia were detected using EVOS2 fluorescence microscopy. B ) Detection of CAT1 and viral protein Env in syncytium. After 48 h incubation with the supernatant of FLK-BLV cells, the transfected CHO-K1 cells were labeled with an anti-gp51 mAb (BLV-1) followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red). Nuclei were stained with Hoechst 33342 (blue). CAT1-EGFP was visualized to determine CAT1 expression (green) in the cells. C ) Ovine, porcine, and bCAT1/SLC7A1 proteins formed syncytia in both FLK-BLV and PK15/pBLV-IF2 cells following BLV infection. The FLK-BLV cells and PK15/pBLV-IF2 cells that stably transfected pBLV-IF2 were transfected with either ovine CAT1/pEGFP-N1, porcine CAT1/pEGFP-N1, bCAT1/pEGFP-N1, or pEGFP-N1 and incubated for 48 h. The cells were then fixed with 3.7% formaldehyde and stained using 10 µg/ml Hoechst 33342. EGFP-expressing syncytia were detected using EVOS2 fluorescence microscopy.

    Journal: The FASEB Journal

    Article Title: CAT1/SLC7A1 acts as a cellular receptor for bovine leukemia virus infection

    doi: 10.1096/fj.201901528R

    Figure Lengend Snippet: CAT1/SLC7A1 did not have species specificity with all demonstrating susceptibility to BLV infection. A ) CAT1/SLC7A1s from 9 different animal species did not have species specificity for BLV infection. The pEGFP-N1–expression plasmids encoding different CAT1/SLC7A1 proteins were derived using various animal species, including cattle (bovine), sheep (ovine), human, monkey (primate), pig (porcine), cat (feline), rat, mouse, and Chinese hamster. The plasmids were constructed and transfected into CHO-K1 cells. After 4 h transfection, the cells were added to culture supernatants of FLK-BLV cells and cultured for 48 h. The cells were then fixed with 3.7% formaldehyde and stained with 10 µg/ml Hoechst 33342. EGFP-expressing syncytia were detected using EVOS2 fluorescence microscopy. B ) Detection of CAT1 and viral protein Env in syncytium. After 48 h incubation with the supernatant of FLK-BLV cells, the transfected CHO-K1 cells were labeled with an anti-gp51 mAb (BLV-1) followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red). Nuclei were stained with Hoechst 33342 (blue). CAT1-EGFP was visualized to determine CAT1 expression (green) in the cells. C ) Ovine, porcine, and bCAT1/SLC7A1 proteins formed syncytia in both FLK-BLV and PK15/pBLV-IF2 cells following BLV infection. The FLK-BLV cells and PK15/pBLV-IF2 cells that stably transfected pBLV-IF2 were transfected with either ovine CAT1/pEGFP-N1, porcine CAT1/pEGFP-N1, bCAT1/pEGFP-N1, or pEGFP-N1 and incubated for 48 h. The cells were then fixed with 3.7% formaldehyde and stained using 10 µg/ml Hoechst 33342. EGFP-expressing syncytia were detected using EVOS2 fluorescence microscopy.

    Article Snippet: Cells were labeled with an anti–BLV Env antibody followed by incubation with Alexa Fluor 594 goat anti-mouse IgG (red fluorescence) to detect cells expressing Env and staining with Hoechst 33342 (blue fluorescence) for detection of the nucleus.

    Techniques: Infection, Expressing, Derivative Assay, Construct, Transfection, Cell Culture, Staining, Fluorescence, Microscopy, Incubation, Labeling, Stable Transfection