fitc-conjugated goat anti-human igg Search Results


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  • 90
    Millipore fitc conjugated goat anti human igg
    Relative binding of HIV-1 broadly neutralizing monoclonal Abs to FcγRI on TZM-bl cells. TZM-bl cells expressing FcγRI were incubated with either IgG1b12, 2G12, 2F5, or 4E10 for 1 h at 4°C. Cells were washed three times, stained with <t>FITC-conjugated</t> goat anti-human <t>IgG</t> (Fab specific), and analyzed by flow cytometry. Top, percentage of cells staining positive; bottom, mean fluorescence intensity (MFI) of positive cells. Background staining with an isotype control Ab was subtracted in both cases.
    Fitc Conjugated Goat Anti Human Igg, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 82 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bio-Rad fitc conjugated goat anti human igg
    Relative binding of HIV-1 broadly neutralizing monoclonal Abs to FcγRI on TZM-bl cells. TZM-bl cells expressing FcγRI were incubated with either IgG1b12, 2G12, 2F5, or 4E10 for 1 h at 4°C. Cells were washed three times, stained with <t>FITC-conjugated</t> goat anti-human <t>IgG</t> (Fab specific), and analyzed by flow cytometry. Top, percentage of cells staining positive; bottom, mean fluorescence intensity (MFI) of positive cells. Background staining with an isotype control Ab was subtracted in both cases.
    Fitc Conjugated Goat Anti Human Igg, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher fitc conjugated goat anti human igg
    E1-G CT colocalizes with proteins involved in generation of ER-derived transport vesicles. BHK-E1-G CT cells were fixed with methanol as described above and incubated with rabbit (A) and mouse (C) antibodies directed against the ERGIC marker p58 or rabbit anti-Sec23p (E). E1-G CT was detected using P5D4 (B), human anti-E1 (D), or rabbit anti-G CT (F). The distribution of Sec23p (G) and mouse anti-G CT (H) in untransfected BHK cells are also shown. Secondary antibodies were <t>FITC-donkey</t> anti-rabbit <t>IgG,</t> FITC-goat anti-human IgG, and Texas Red-goat anti-mouse IgG. Overlap between p58 and Sec23p with E1-G CT in the juxtanuclear region is indicated by arrowheads. Bar, 10 μm.
    Fitc Conjugated Goat Anti Human Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam fitc conjugated goat anti human igg
    Functional evaluation of the recombinant anti-CD20 mAb in vitro . ( a , b ) CD20-binding activity of the recombinant mAb. Daudi ( a ) and Raji ( b ) cells were incubated with serial log dilutions of the mAbs, and their binding activities were assayed by flow cytometry using a <t>FITC-conjugated</t> goat anti-human <t>IgG</t> (H + L). Data are presented as the mean ± SD (n = 3). ( c , d ) CDC induced by the anti-CD20 mAb. Daudi ( c ) and Raji ( d ) cells were incubated with increasing concentrations of mAbs in the presence of human serum. CDC activity was measured using a Luminescent Cell Viability Assay Kit. Data are presented as the mean ± SD (n = 3). ( e ) ADCC induced by the anti-CD20 mAb. ADCC activity against Raji cells was determined using human PBMCs as effector cells at an effector-to-target cell ratio of 5:1. The activity was measured using a standard LDH assay. Data are presented as the mean ± SD (n = 3).
    Fitc Conjugated Goat Anti Human Igg, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology fitc conjugated goat anti human igg
    Functional evaluation of the recombinant anti-CD20 mAb in vitro . ( a , b ) CD20-binding activity of the recombinant mAb. Daudi ( a ) and Raji ( b ) cells were incubated with serial log dilutions of the mAbs, and their binding activities were assayed by flow cytometry using a <t>FITC-conjugated</t> goat anti-human <t>IgG</t> (H + L). Data are presented as the mean ± SD (n = 3). ( c , d ) CDC induced by the anti-CD20 mAb. Daudi ( c ) and Raji ( d ) cells were incubated with increasing concentrations of mAbs in the presence of human serum. CDC activity was measured using a Luminescent Cell Viability Assay Kit. Data are presented as the mean ± SD (n = 3). ( e ) ADCC induced by the anti-CD20 mAb. ADCC activity against Raji cells was determined using human PBMCs as effector cells at an effector-to-target cell ratio of 5:1. The activity was measured using a standard LDH assay. Data are presented as the mean ± SD (n = 3).
    Fitc Conjugated Goat Anti Human Igg, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Immunotec fitc conjugated goat anti human igg
    Functional evaluation of the recombinant anti-CD20 mAb in vitro . ( a , b ) CD20-binding activity of the recombinant mAb. Daudi ( a ) and Raji ( b ) cells were incubated with serial log dilutions of the mAbs, and their binding activities were assayed by flow cytometry using a <t>FITC-conjugated</t> goat anti-human <t>IgG</t> (H + L). Data are presented as the mean ± SD (n = 3). ( c , d ) CDC induced by the anti-CD20 mAb. Daudi ( c ) and Raji ( d ) cells were incubated with increasing concentrations of mAbs in the presence of human serum. CDC activity was measured using a Luminescent Cell Viability Assay Kit. Data are presented as the mean ± SD (n = 3). ( e ) ADCC induced by the anti-CD20 mAb. ADCC activity against Raji cells was determined using human PBMCs as effector cells at an effector-to-target cell ratio of 5:1. The activity was measured using a standard LDH assay. Data are presented as the mean ± SD (n = 3).
    Fitc Conjugated Goat Anti Human Igg, supplied by Immunotec, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Proteintech fitc conjugated goat anti human igg
    Autoantibody production and immune cell infiltration in the AIH mouse model. a Representative pictures of rat liver sections stained with plasma from AIH mice (primary antibody) at day 35, followed by Alexa Fluor 488-conjugated anti-mouse <t>IgG</t> (secondary antibody). The plasma from AIH-1 (type-1 autoimmune hepatitis) patients (antinuclear antibodies, ANAs), AIH-2 (type-2 autoimmune hepatitis) patients (anti-liver-kidney microsomal-1 (LKM-1) antibodies), and PBC patients (anti-mitochondrial antibodies, AMAs) were used to stain the same rat liver sections as the primary antibodies. <t>FITC-conjugated</t> anti-human IgG was used as the secondary antibody (n = 3, upper column: ×200 magnification, lower column: ×800 magnification). b The dilution ratio of autoantibodies in the plasma of AIH mouse after the first injection (n = 7). c CD4 + T cells and CD8 + T cells were found to accumulate in the AIH mouse liver on different days after the first injection, with CD4 (red) and CD8 (green) as their marker respectively. Nuclei were stained with Hoechst (blue) (n = 5, ×600 magnification). **P
    Fitc Conjugated Goat Anti Human Igg, supplied by Proteintech, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Beckman Coulter fitc conjugated goat anti human igg ab
    Autoantibody production and immune cell infiltration in the AIH mouse model. a Representative pictures of rat liver sections stained with plasma from AIH mice (primary antibody) at day 35, followed by Alexa Fluor 488-conjugated anti-mouse <t>IgG</t> (secondary antibody). The plasma from AIH-1 (type-1 autoimmune hepatitis) patients (antinuclear antibodies, ANAs), AIH-2 (type-2 autoimmune hepatitis) patients (anti-liver-kidney microsomal-1 (LKM-1) antibodies), and PBC patients (anti-mitochondrial antibodies, AMAs) were used to stain the same rat liver sections as the primary antibodies. <t>FITC-conjugated</t> anti-human IgG was used as the secondary antibody (n = 3, upper column: ×200 magnification, lower column: ×800 magnification). b The dilution ratio of autoantibodies in the plasma of AIH mouse after the first injection (n = 7). c CD4 + T cells and CD8 + T cells were found to accumulate in the AIH mouse liver on different days after the first injection, with CD4 (red) and CD8 (green) as their marker respectively. Nuclei were stained with Hoechst (blue) (n = 5, ×600 magnification). **P
    Fitc Conjugated Goat Anti Human Igg Ab, supplied by Beckman Coulter, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ICN Biomedicals fitc conjugated goat anti human igg
    Autoantibody production and immune cell infiltration in the AIH mouse model. a Representative pictures of rat liver sections stained with plasma from AIH mice (primary antibody) at day 35, followed by Alexa Fluor 488-conjugated anti-mouse <t>IgG</t> (secondary antibody). The plasma from AIH-1 (type-1 autoimmune hepatitis) patients (antinuclear antibodies, ANAs), AIH-2 (type-2 autoimmune hepatitis) patients (anti-liver-kidney microsomal-1 (LKM-1) antibodies), and PBC patients (anti-mitochondrial antibodies, AMAs) were used to stain the same rat liver sections as the primary antibodies. <t>FITC-conjugated</t> anti-human IgG was used as the secondary antibody (n = 3, upper column: ×200 magnification, lower column: ×800 magnification). b The dilution ratio of autoantibodies in the plasma of AIH mouse after the first injection (n = 7). c CD4 + T cells and CD8 + T cells were found to accumulate in the AIH mouse liver on different days after the first injection, with CD4 (red) and CD8 (green) as their marker respectively. Nuclei were stained with Hoechst (blue) (n = 5, ×600 magnification). **P
    Fitc Conjugated Goat Anti Human Igg, supplied by ICN Biomedicals, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Jackson Immuno goat anti human fitc conjugated igg
    In vitro activation and allelic exclusion. ( A ) Upregulation of CTLA4 binding following a 24-h culture with antiIgM or medium alone ( Ctrl ). Cultured splenocytes from transgenic mice in a μMT background (or wild-type nontransgenic controls, WT, Non-tg) were stained with PE–anti-B220 and an mCTLA4–Hγ1 fusion protein, which was visualized with <t>FITC–anti-human</t> <t>IgG.</t> The profiles were gated for B220 + cells that excluded propidium iodide. ( B ) Exclusion of endogenous IgD. Splenocytes were stained with FITC–antiB220, PE–anti-IgD, and biotinylated anti-D1.3 plus RED670– streptavidin. Cells were gated for B220 + and numbers denote percentage of splenic B cells within each quadrant.
    Goat Anti Human Fitc Conjugated Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ZEUS Scientific fitc conjugated goat anti human igg
    In vitro activation and allelic exclusion. ( A ) Upregulation of CTLA4 binding following a 24-h culture with antiIgM or medium alone ( Ctrl ). Cultured splenocytes from transgenic mice in a μMT background (or wild-type nontransgenic controls, WT, Non-tg) were stained with PE–anti-B220 and an mCTLA4–Hγ1 fusion protein, which was visualized with <t>FITC–anti-human</t> <t>IgG.</t> The profiles were gated for B220 + cells that excluded propidium iodide. ( B ) Exclusion of endogenous IgD. Splenocytes were stained with FITC–antiB220, PE–anti-IgD, and biotinylated anti-D1.3 plus RED670– streptavidin. Cells were gated for B220 + and numbers denote percentage of splenic B cells within each quadrant.
    Fitc Conjugated Goat Anti Human Igg, supplied by ZEUS Scientific, used in various techniques. Bioz Stars score: 90/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Jackson Laboratory fitc conjugated goat anti human igg ab
    In vitro activation and allelic exclusion. ( A ) Upregulation of CTLA4 binding following a 24-h culture with antiIgM or medium alone ( Ctrl ). Cultured splenocytes from transgenic mice in a μMT background (or wild-type nontransgenic controls, WT, Non-tg) were stained with PE–anti-B220 and an mCTLA4–Hγ1 fusion protein, which was visualized with <t>FITC–anti-human</t> <t>IgG.</t> The profiles were gated for B220 + cells that excluded propidium iodide. ( B ) Exclusion of endogenous IgD. Splenocytes were stained with FITC–antiB220, PE–anti-IgD, and biotinylated anti-D1.3 plus RED670– streptavidin. Cells were gated for B220 + and numbers denote percentage of splenic B cells within each quadrant.
    Fitc Conjugated Goat Anti Human Igg Ab, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 89/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Sihuan fitc conjugated goat anti human igg
    In vitro activation and allelic exclusion. ( A ) Upregulation of CTLA4 binding following a 24-h culture with antiIgM or medium alone ( Ctrl ). Cultured splenocytes from transgenic mice in a μMT background (or wild-type nontransgenic controls, WT, Non-tg) were stained with PE–anti-B220 and an mCTLA4–Hγ1 fusion protein, which was visualized with <t>FITC–anti-human</t> <t>IgG.</t> The profiles were gated for B220 + cells that excluded propidium iodide. ( B ) Exclusion of endogenous IgD. Splenocytes were stained with FITC–antiB220, PE–anti-IgD, and biotinylated anti-D1.3 plus RED670– streptavidin. Cells were gated for B220 + and numbers denote percentage of splenic B cells within each quadrant.
    Fitc Conjugated Goat Anti Human Igg, supplied by Sihuan, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher goat anti human igg fitc conjugates
    IL-1β secretion induced by O. tsutsugamushi infection in LPS-primed BMDMs was blocked by pan-caspase inhibitor. LPS-primed BMDMs were pretreated with pan-caspase inhibitor Z-VAD-FMK (VAD) at the indicated concentration for 1 h and then challenged with ATP (5 mM) for 3 h or OT (A. ICU/cell=10; B,C. ICU/cell=50) for the indicated period (A) or 6 h (B, C). (A) Cells were fixed and stained with human antiserum against OT and a <t>FITC-conjugated</t> goat anti-human <t>IgG</t> antibody, and examined using a fluorescence microscope. Green spots indicate OT and red areas indicate the host cell. Scale bar, 50 µm. (B) Cell viability was measured by MTT assay. (C) The production of IL-1β and IL-6 was assessed by ELISA. Error bars represent SD of triplicate samples. **p
    Goat Anti Human Igg Fitc Conjugates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    SouthernBiotech fitc conjugated goat anti human igg
    Immunofluorescence reactivity of <t>IgG</t> from patient sera on LoVo cell line ADAM10 pro-domain: reactivity of the rabbit polyclonal anti-ADAM10 pro-domain-specific Ab (ab39178, Abcam), which recognized the immature non-functional isoform of ADAM10, showed patched reactivity. ADAM10 ectodomain: reactivity of the goat polyclonal anti-ADAM10 ectodomain-specific Ab (R D-Systems, AB936), which recognized both the immature and mature (cleaved and functional) isoforms of ADAM10, showed patched and diffuse signals. Anti-ADAM10 auto-Ab positive serum: reactivity of the human IgG fraction purified from a representative serum (C2) of Crc patients considered positive for the presence of auto-Ab anti ADAM10, showed patched reactivity. Anti-ADAM10 auto-Ab negative serum: no reactivity was observed for the human IgG fraction purified from serum (C25) of a representative Crc patient considered negative for the presence of auto-Ab anti ADAM10. Cell nuclei were stained with Hoechst-33342. The reactivity of the secondary antibodies (goat anti-rabbit IgG Alexa-488; donkey anti-goat IgG Alexa-488 and goat anti-human IgG <t>FITC)</t> are shown as negative controls in SM-Figure 3 . Images were acquired by immunofluorescence microscopy (Zeiss Upright Axo Imager 2 equipped with AxoVision Rel.4.8.2 software); magnification 63X. Images were linearly adjusted for brightness and contrast using Adobe-Photoshop CS4 v.11 software.
    Fitc Conjugated Goat Anti Human Igg, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 93/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Jackson Immuno fitc conjugated affinipure goat anti human igg
    SARS-CoV RBD spike protein was internalized by the susceptible cells together with ACE2. (A) Decreased cell surface-bound RBD-Fc protein following incubation with VeroE6 cells at 37 °C compared to 4 °C. After incubation with VeroE6 cells for 3 h, cell surface-bound Fc or RBD-Fc proteins were detected using a <t>FITC-conjugated</t> human <t>IgG-specific</t> antibody. Representative FACS histograms are shown for three independent experiments. Blue and black histograms represent the results from the cells incubated with the RBD-Fc or Fc at 4 °C, respectively. The red histogram represents the results from the cells incubated with the RBD-Fc at 37 °C. (B) ACE2 internalization is the RBD spike protein-dependent. After ACE2-GFP-293 cells were incubated with the RBD-Fc or Fc protein at 37 °C for 3 h, the distribution of ACE2-GFP molecules in live cells was observed under a fluorescence microscope. (C) Co-localization of ACE2 and the RBD of SARS-CoV in ACE2-GFP-293 cells under a confocal laser-scanning microscope. After incubation with the RBD-Fc or Fc protein at 37 °C for 3 h, ACE2-GFP-293 cells were washed to remove excess RBD-Fc or Fc protein and then fixed with 4% paraformaldehyde. Immunostaining was performed using an Alexa Fluor 568-conjugated human IgG-specific antibody to detect the distribution of the Fc or RBD-Fc protein in the cells. (D) and (E) Statistical analysis of the differences between the RBD-Fc and Fc protein treatments (Chi-square test). ** P
    Fitc Conjugated Affinipure Goat Anti Human Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    SouthernBiotech goat anti human igg fluorescein isothiocyanate fitc conjugate
    SARS-CoV RBD spike protein was internalized by the susceptible cells together with ACE2. (A) Decreased cell surface-bound RBD-Fc protein following incubation with VeroE6 cells at 37 °C compared to 4 °C. After incubation with VeroE6 cells for 3 h, cell surface-bound Fc or RBD-Fc proteins were detected using a <t>FITC-conjugated</t> human <t>IgG-specific</t> antibody. Representative FACS histograms are shown for three independent experiments. Blue and black histograms represent the results from the cells incubated with the RBD-Fc or Fc at 4 °C, respectively. The red histogram represents the results from the cells incubated with the RBD-Fc at 37 °C. (B) ACE2 internalization is the RBD spike protein-dependent. After ACE2-GFP-293 cells were incubated with the RBD-Fc or Fc protein at 37 °C for 3 h, the distribution of ACE2-GFP molecules in live cells was observed under a fluorescence microscope. (C) Co-localization of ACE2 and the RBD of SARS-CoV in ACE2-GFP-293 cells under a confocal laser-scanning microscope. After incubation with the RBD-Fc or Fc protein at 37 °C for 3 h, ACE2-GFP-293 cells were washed to remove excess RBD-Fc or Fc protein and then fixed with 4% paraformaldehyde. Immunostaining was performed using an Alexa Fluor 568-conjugated human IgG-specific antibody to detect the distribution of the Fc or RBD-Fc protein in the cells. (D) and (E) Statistical analysis of the differences between the RBD-Fc and Fc protein treatments (Chi-square test). ** P
    Goat Anti Human Igg Fluorescein Isothiocyanate Fitc Conjugate, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Kent Laboratories polyclonal fitc conjugated goat anti human igg
    Colocalization of <t>IgG</t> and LRP2. (A–C) Immunofluorescence experiments performed on cryosections of normal human kidney tissue. (A) Staining for LRP2 (red) using a rabbit <t>polyclonal</t> antibody along the apical membrane of the tubular epithelium. (B) Indirect immunofluorescence of serum from a patient with ABBA on the normal human kidney. (C) Strong colocalization of LRP2 and serum antibodies from a patient with ABBA. (D–F) Immunofluorescence experiments performed on a renal biopsy sample from a patient with anti-LRP2 nephropathy shows granular TBM staining in a proximal tubule for LRP2 using (D) a mouse mAb and (E) IgG. (F) There is strong colocalization of LRP2 and IgG in the TBM deposits. (G–I) Immunofluorescence experiments performed on a renal biopsy sample from a patient with lupus nephritis shows staining of the tubular apical membrane for LRP2 using (G) a mouse mAb and (H) granular TBM staining for IgG. (I) There is complete lack of colocalization in a patient without anti-LRP2 nephropathy.
    Polyclonal Fitc Conjugated Goat Anti Human Igg, supplied by Kent Laboratories, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Relative binding of HIV-1 broadly neutralizing monoclonal Abs to FcγRI on TZM-bl cells. TZM-bl cells expressing FcγRI were incubated with either IgG1b12, 2G12, 2F5, or 4E10 for 1 h at 4°C. Cells were washed three times, stained with FITC-conjugated goat anti-human IgG (Fab specific), and analyzed by flow cytometry. Top, percentage of cells staining positive; bottom, mean fluorescence intensity (MFI) of positive cells. Background staining with an isotype control Ab was subtracted in both cases.

    Journal: Journal of Virology

    Article Title: Utilization of Immunoglobulin G Fc Receptors by Human Immunodeficiency Virus Type 1: a Specific Role for Antibodies against the Membrane-Proximal External Region of gp41 ▿

    doi: 10.1128/JVI.00656-09

    Figure Lengend Snippet: Relative binding of HIV-1 broadly neutralizing monoclonal Abs to FcγRI on TZM-bl cells. TZM-bl cells expressing FcγRI were incubated with either IgG1b12, 2G12, 2F5, or 4E10 for 1 h at 4°C. Cells were washed three times, stained with FITC-conjugated goat anti-human IgG (Fab specific), and analyzed by flow cytometry. Top, percentage of cells staining positive; bottom, mean fluorescence intensity (MFI) of positive cells. Background staining with an isotype control Ab was subtracted in both cases.

    Article Snippet: Binding was quantified by flow cytometry by using FITC-conjugated goat anti-human IgG (Fab specific; Sigma-Aldrich, St. Louis, MO).

    Techniques: Binding Assay, Expressing, Incubation, Staining, Flow Cytometry, Cytometry, Fluorescence

    E1-G CT colocalizes with proteins involved in generation of ER-derived transport vesicles. BHK-E1-G CT cells were fixed with methanol as described above and incubated with rabbit (A) and mouse (C) antibodies directed against the ERGIC marker p58 or rabbit anti-Sec23p (E). E1-G CT was detected using P5D4 (B), human anti-E1 (D), or rabbit anti-G CT (F). The distribution of Sec23p (G) and mouse anti-G CT (H) in untransfected BHK cells are also shown. Secondary antibodies were FITC-donkey anti-rabbit IgG, FITC-goat anti-human IgG, and Texas Red-goat anti-mouse IgG. Overlap between p58 and Sec23p with E1-G CT in the juxtanuclear region is indicated by arrowheads. Bar, 10 μm.

    Journal: Molecular Biology of the Cell

    Article Title: Immunoisolation and Characterization of a Subdomain of the Endoplasmic Reticulum That Concentrates Proteins Involved in COPII Vesicle Biogenesis

    doi:

    Figure Lengend Snippet: E1-G CT colocalizes with proteins involved in generation of ER-derived transport vesicles. BHK-E1-G CT cells were fixed with methanol as described above and incubated with rabbit (A) and mouse (C) antibodies directed against the ERGIC marker p58 or rabbit anti-Sec23p (E). E1-G CT was detected using P5D4 (B), human anti-E1 (D), or rabbit anti-G CT (F). The distribution of Sec23p (G) and mouse anti-G CT (H) in untransfected BHK cells are also shown. Secondary antibodies were FITC-donkey anti-rabbit IgG, FITC-goat anti-human IgG, and Texas Red-goat anti-mouse IgG. Overlap between p58 and Sec23p with E1-G CT in the juxtanuclear region is indicated by arrowheads. Bar, 10 μm.

    Article Snippet: FITC-conjugated goat anti-human IgG was purchased from Zymed (South San Francisco, CA).

    Techniques: Derivative Assay, Incubation, Marker

    E1-G CT does not colocalize with Golgi or RER markers in BHK cells. Stably transfected BHK cells expressing E1-G CT grown on 12-mm coverslips were fixed and permeabilized with methanol at −20°C before processing for double label indirect immunofluorescence. Golgi and RER membranes were labeled, respectively, with rabbit anti-mannosidase II (Man II) and anticalnexin in panels A and C, respectively. E1-G CT was stained in panels B and D using a monoclonal antibody, P5D4, directed against the VSV G CT domain. Secondary antibodies were FITC-donkey anti-rabbit IgG and Texas Red-goat anti-mouse IgG. Bar, 10 μm.

    Journal: Molecular Biology of the Cell

    Article Title: Immunoisolation and Characterization of a Subdomain of the Endoplasmic Reticulum That Concentrates Proteins Involved in COPII Vesicle Biogenesis

    doi:

    Figure Lengend Snippet: E1-G CT does not colocalize with Golgi or RER markers in BHK cells. Stably transfected BHK cells expressing E1-G CT grown on 12-mm coverslips were fixed and permeabilized with methanol at −20°C before processing for double label indirect immunofluorescence. Golgi and RER membranes were labeled, respectively, with rabbit anti-mannosidase II (Man II) and anticalnexin in panels A and C, respectively. E1-G CT was stained in panels B and D using a monoclonal antibody, P5D4, directed against the VSV G CT domain. Secondary antibodies were FITC-donkey anti-rabbit IgG and Texas Red-goat anti-mouse IgG. Bar, 10 μm.

    Article Snippet: FITC-conjugated goat anti-human IgG was purchased from Zymed (South San Francisco, CA).

    Techniques: Stable Transfection, Transfection, Expressing, Immunofluorescence, Labeling, Staining

    Functional evaluation of the recombinant anti-CD20 mAb in vitro . ( a , b ) CD20-binding activity of the recombinant mAb. Daudi ( a ) and Raji ( b ) cells were incubated with serial log dilutions of the mAbs, and their binding activities were assayed by flow cytometry using a FITC-conjugated goat anti-human IgG (H + L). Data are presented as the mean ± SD (n = 3). ( c , d ) CDC induced by the anti-CD20 mAb. Daudi ( c ) and Raji ( d ) cells were incubated with increasing concentrations of mAbs in the presence of human serum. CDC activity was measured using a Luminescent Cell Viability Assay Kit. Data are presented as the mean ± SD (n = 3). ( e ) ADCC induced by the anti-CD20 mAb. ADCC activity against Raji cells was determined using human PBMCs as effector cells at an effector-to-target cell ratio of 5:1. The activity was measured using a standard LDH assay. Data are presented as the mean ± SD (n = 3).

    Journal: Scientific Reports

    Article Title: A novel glycosylated anti-CD20 monoclonal antibody from transgenic cattle

    doi: 10.1038/s41598-018-31417-2

    Figure Lengend Snippet: Functional evaluation of the recombinant anti-CD20 mAb in vitro . ( a , b ) CD20-binding activity of the recombinant mAb. Daudi ( a ) and Raji ( b ) cells were incubated with serial log dilutions of the mAbs, and their binding activities were assayed by flow cytometry using a FITC-conjugated goat anti-human IgG (H + L). Data are presented as the mean ± SD (n = 3). ( c , d ) CDC induced by the anti-CD20 mAb. Daudi ( c ) and Raji ( d ) cells were incubated with increasing concentrations of mAbs in the presence of human serum. CDC activity was measured using a Luminescent Cell Viability Assay Kit. Data are presented as the mean ± SD (n = 3). ( e ) ADCC induced by the anti-CD20 mAb. ADCC activity against Raji cells was determined using human PBMCs as effector cells at an effector-to-target cell ratio of 5:1. The activity was measured using a standard LDH assay. Data are presented as the mean ± SD (n = 3).

    Article Snippet: Binding assay Daudi and Raji cells were incubated with serially diluted anti-CD20 mAb at 37 °C for 1 h. The cells were incubated with a FITC-conjugated goat anti-human IgG (H + L) (Abcam, Cambridge, MA) for 1 h at 4 °C and analysed with a FACSCalibur system (BD Biosciences, San Jose, CA).

    Techniques: Functional Assay, Recombinant, In Vitro, Binding Assay, Activity Assay, Incubation, Flow Cytometry, Cytometry, Cell Viability Assay, Lactate Dehydrogenase Assay

    Autoantibody production and immune cell infiltration in the AIH mouse model. a Representative pictures of rat liver sections stained with plasma from AIH mice (primary antibody) at day 35, followed by Alexa Fluor 488-conjugated anti-mouse IgG (secondary antibody). The plasma from AIH-1 (type-1 autoimmune hepatitis) patients (antinuclear antibodies, ANAs), AIH-2 (type-2 autoimmune hepatitis) patients (anti-liver-kidney microsomal-1 (LKM-1) antibodies), and PBC patients (anti-mitochondrial antibodies, AMAs) were used to stain the same rat liver sections as the primary antibodies. FITC-conjugated anti-human IgG was used as the secondary antibody (n = 3, upper column: ×200 magnification, lower column: ×800 magnification). b The dilution ratio of autoantibodies in the plasma of AIH mouse after the first injection (n = 7). c CD4 + T cells and CD8 + T cells were found to accumulate in the AIH mouse liver on different days after the first injection, with CD4 (red) and CD8 (green) as their marker respectively. Nuclei were stained with Hoechst (blue) (n = 5, ×600 magnification). **P

    Journal: Journal of Translational Medicine

    Article Title: Plasma proteomic analysis of autoimmune hepatitis in an improved AIH mouse model

    doi: 10.1186/s12967-019-02180-3

    Figure Lengend Snippet: Autoantibody production and immune cell infiltration in the AIH mouse model. a Representative pictures of rat liver sections stained with plasma from AIH mice (primary antibody) at day 35, followed by Alexa Fluor 488-conjugated anti-mouse IgG (secondary antibody). The plasma from AIH-1 (type-1 autoimmune hepatitis) patients (antinuclear antibodies, ANAs), AIH-2 (type-2 autoimmune hepatitis) patients (anti-liver-kidney microsomal-1 (LKM-1) antibodies), and PBC patients (anti-mitochondrial antibodies, AMAs) were used to stain the same rat liver sections as the primary antibodies. FITC-conjugated anti-human IgG was used as the secondary antibody (n = 3, upper column: ×200 magnification, lower column: ×800 magnification). b The dilution ratio of autoantibodies in the plasma of AIH mouse after the first injection (n = 7). c CD4 + T cells and CD8 + T cells were found to accumulate in the AIH mouse liver on different days after the first injection, with CD4 (red) and CD8 (green) as their marker respectively. Nuclei were stained with Hoechst (blue) (n = 5, ×600 magnification). **P

    Article Snippet: The plasma from AIH mice and patients were diluted 1:300 and used as the primary antibody and then detected using DyLight 488 conjugated goat anti-mouse IgG (1:100; A23210, Abbkine, Waltham, MA USA) and FITC-conjugated goat anti-human IgG (1:100; SA00003-12, Proteintech, Rosemont, IL USA).

    Techniques: Staining, Mouse Assay, Injection, Marker

    In vitro activation and allelic exclusion. ( A ) Upregulation of CTLA4 binding following a 24-h culture with antiIgM or medium alone ( Ctrl ). Cultured splenocytes from transgenic mice in a μMT background (or wild-type nontransgenic controls, WT, Non-tg) were stained with PE–anti-B220 and an mCTLA4–Hγ1 fusion protein, which was visualized with FITC–anti-human IgG. The profiles were gated for B220 + cells that excluded propidium iodide. ( B ) Exclusion of endogenous IgD. Splenocytes were stained with FITC–antiB220, PE–anti-IgD, and biotinylated anti-D1.3 plus RED670– streptavidin. Cells were gated for B220 + and numbers denote percentage of splenic B cells within each quadrant.

    Journal: The Journal of Experimental Medicine

    Article Title: The Immunoglobulin (Ig)? and Ig? Cytoplasmic Domains Are Independently Sufficient to Signal B Cell Maturation and Activation in Transgenic Mice

    doi:

    Figure Lengend Snippet: In vitro activation and allelic exclusion. ( A ) Upregulation of CTLA4 binding following a 24-h culture with antiIgM or medium alone ( Ctrl ). Cultured splenocytes from transgenic mice in a μMT background (or wild-type nontransgenic controls, WT, Non-tg) were stained with PE–anti-B220 and an mCTLA4–Hγ1 fusion protein, which was visualized with FITC–anti-human IgG. The profiles were gated for B220 + cells that excluded propidium iodide. ( B ) Exclusion of endogenous IgD. Splenocytes were stained with FITC–antiB220, PE–anti-IgD, and biotinylated anti-D1.3 plus RED670– streptavidin. Cells were gated for B220 + and numbers denote percentage of splenic B cells within each quadrant.

    Article Snippet: To monitor CTLA4 binding, spleen cells depleted of erythrocytes were cultured (106 cells/ml) in medium in the presence or absence of 10 μg/ml F(ab′)2 goat anti–mouse IgM (Jackson ImmunoResearch) for 24 h before staining with mCTLA4–Hγ1 fusion protein (gift from P. Lane) and FITC-conjugated goat anti– human IgG (Jackson ImmunoResearch).

    Techniques: In Vitro, Activation Assay, Binding Assay, Cell Culture, Transgenic Assay, Mouse Assay, Staining

    IL-1β secretion induced by O. tsutsugamushi infection in LPS-primed BMDMs was blocked by pan-caspase inhibitor. LPS-primed BMDMs were pretreated with pan-caspase inhibitor Z-VAD-FMK (VAD) at the indicated concentration for 1 h and then challenged with ATP (5 mM) for 3 h or OT (A. ICU/cell=10; B,C. ICU/cell=50) for the indicated period (A) or 6 h (B, C). (A) Cells were fixed and stained with human antiserum against OT and a FITC-conjugated goat anti-human IgG antibody, and examined using a fluorescence microscope. Green spots indicate OT and red areas indicate the host cell. Scale bar, 50 µm. (B) Cell viability was measured by MTT assay. (C) The production of IL-1β and IL-6 was assessed by ELISA. Error bars represent SD of triplicate samples. **p

    Journal: PLoS ONE

    Article Title: Intracellular Invasion of Orientia tsutsugamushi Activates Inflammasome in ASC-Dependent Manner

    doi: 10.1371/journal.pone.0039042

    Figure Lengend Snippet: IL-1β secretion induced by O. tsutsugamushi infection in LPS-primed BMDMs was blocked by pan-caspase inhibitor. LPS-primed BMDMs were pretreated with pan-caspase inhibitor Z-VAD-FMK (VAD) at the indicated concentration for 1 h and then challenged with ATP (5 mM) for 3 h or OT (A. ICU/cell=10; B,C. ICU/cell=50) for the indicated period (A) or 6 h (B, C). (A) Cells were fixed and stained with human antiserum against OT and a FITC-conjugated goat anti-human IgG antibody, and examined using a fluorescence microscope. Green spots indicate OT and red areas indicate the host cell. Scale bar, 50 µm. (B) Cell viability was measured by MTT assay. (C) The production of IL-1β and IL-6 was assessed by ELISA. Error bars represent SD of triplicate samples. **p

    Article Snippet: After the slide was dried, it was stained with goat anti-human IgG-FITC conjugates (Caltag, CA, USA) and incubated in the dark at 37°C for 30 min.

    Techniques: Infection, Concentration Assay, Staining, Fluorescence, Microscopy, MTT Assay, Enzyme-linked Immunosorbent Assay

    Immunofluorescence reactivity of IgG from patient sera on LoVo cell line ADAM10 pro-domain: reactivity of the rabbit polyclonal anti-ADAM10 pro-domain-specific Ab (ab39178, Abcam), which recognized the immature non-functional isoform of ADAM10, showed patched reactivity. ADAM10 ectodomain: reactivity of the goat polyclonal anti-ADAM10 ectodomain-specific Ab (R D-Systems, AB936), which recognized both the immature and mature (cleaved and functional) isoforms of ADAM10, showed patched and diffuse signals. Anti-ADAM10 auto-Ab positive serum: reactivity of the human IgG fraction purified from a representative serum (C2) of Crc patients considered positive for the presence of auto-Ab anti ADAM10, showed patched reactivity. Anti-ADAM10 auto-Ab negative serum: no reactivity was observed for the human IgG fraction purified from serum (C25) of a representative Crc patient considered negative for the presence of auto-Ab anti ADAM10. Cell nuclei were stained with Hoechst-33342. The reactivity of the secondary antibodies (goat anti-rabbit IgG Alexa-488; donkey anti-goat IgG Alexa-488 and goat anti-human IgG FITC) are shown as negative controls in SM-Figure 3 . Images were acquired by immunofluorescence microscopy (Zeiss Upright Axo Imager 2 equipped with AxoVision Rel.4.8.2 software); magnification 63X. Images were linearly adjusted for brightness and contrast using Adobe-Photoshop CS4 v.11 software.

    Journal: Oncotarget

    Article Title: Serological immune response against ADAM10 pro-domain is associated with favourable prognosis in stage III colorectal cancer patients

    doi: 10.18632/oncotarget.11181

    Figure Lengend Snippet: Immunofluorescence reactivity of IgG from patient sera on LoVo cell line ADAM10 pro-domain: reactivity of the rabbit polyclonal anti-ADAM10 pro-domain-specific Ab (ab39178, Abcam), which recognized the immature non-functional isoform of ADAM10, showed patched reactivity. ADAM10 ectodomain: reactivity of the goat polyclonal anti-ADAM10 ectodomain-specific Ab (R D-Systems, AB936), which recognized both the immature and mature (cleaved and functional) isoforms of ADAM10, showed patched and diffuse signals. Anti-ADAM10 auto-Ab positive serum: reactivity of the human IgG fraction purified from a representative serum (C2) of Crc patients considered positive for the presence of auto-Ab anti ADAM10, showed patched reactivity. Anti-ADAM10 auto-Ab negative serum: no reactivity was observed for the human IgG fraction purified from serum (C25) of a representative Crc patient considered negative for the presence of auto-Ab anti ADAM10. Cell nuclei were stained with Hoechst-33342. The reactivity of the secondary antibodies (goat anti-rabbit IgG Alexa-488; donkey anti-goat IgG Alexa-488 and goat anti-human IgG FITC) are shown as negative controls in SM-Figure 3 . Images were acquired by immunofluorescence microscopy (Zeiss Upright Axo Imager 2 equipped with AxoVision Rel.4.8.2 software); magnification 63X. Images were linearly adjusted for brightness and contrast using Adobe-Photoshop CS4 v.11 software.

    Article Snippet: Alexa-488-conjugated goat-anti-rabbit IgG, donkey-anti-goat IgG and rabbit-anti-mouse IgG (Invitrogen) or FITC-conjugated goat-anti-human IgG (Southern Biotechnology) were used as secondary Abs; in double staining experiments the Alexa-546-conjugated goat-anti-rabbit IgG (Invitrogen) was used.

    Techniques: Immunofluorescence, Functional Assay, Purification, Staining, Microscopy, Software

    Immunofluorescence reactivity after competition with anti ADAM10 pro-domain Ab Anti-ADAM10 pro-domain: reactivity of the rabbit anti-ADAM10 pro-domain Ab (ab39178, Abcam) after competition with control rabbit IgG. Anti-ADAM10 positive serum (C2): reactivity of the IgG fraction purified from a representative serum of a Crc patient considered positive for the presence of auto-Abs anti ADAM10 after competition either with control rabbit IgGs or the anti-ADAM10 pro-domain Ab. Anti-ADAM10 negative serum (C32): reactivity of the IgG fraction purified from a representative serum of a Crc patient considered negative for the presence of anti ADAM10 auto-Ab after competition either with control rabbit IgGs or anti-ADAM10 pro-domain Ab. Cell nuclei were stained with Hoechst-33342; secondary Abs were goat anti-rabbit IgG Alexa-488 and goat anti-human IgG FITC. Images were acquired by immunofluorescence microscopy (Zeiss Upright Axo Imager 2 equipped with AxoVision Rel.4.8.2 software); magnification 63X. Images were linearly adjusted for brightness and contrast using Adobe-Photoshop CS4 v.11 software.

    Journal: Oncotarget

    Article Title: Serological immune response against ADAM10 pro-domain is associated with favourable prognosis in stage III colorectal cancer patients

    doi: 10.18632/oncotarget.11181

    Figure Lengend Snippet: Immunofluorescence reactivity after competition with anti ADAM10 pro-domain Ab Anti-ADAM10 pro-domain: reactivity of the rabbit anti-ADAM10 pro-domain Ab (ab39178, Abcam) after competition with control rabbit IgG. Anti-ADAM10 positive serum (C2): reactivity of the IgG fraction purified from a representative serum of a Crc patient considered positive for the presence of auto-Abs anti ADAM10 after competition either with control rabbit IgGs or the anti-ADAM10 pro-domain Ab. Anti-ADAM10 negative serum (C32): reactivity of the IgG fraction purified from a representative serum of a Crc patient considered negative for the presence of anti ADAM10 auto-Ab after competition either with control rabbit IgGs or anti-ADAM10 pro-domain Ab. Cell nuclei were stained with Hoechst-33342; secondary Abs were goat anti-rabbit IgG Alexa-488 and goat anti-human IgG FITC. Images were acquired by immunofluorescence microscopy (Zeiss Upright Axo Imager 2 equipped with AxoVision Rel.4.8.2 software); magnification 63X. Images were linearly adjusted for brightness and contrast using Adobe-Photoshop CS4 v.11 software.

    Article Snippet: Alexa-488-conjugated goat-anti-rabbit IgG, donkey-anti-goat IgG and rabbit-anti-mouse IgG (Invitrogen) or FITC-conjugated goat-anti-human IgG (Southern Biotechnology) were used as secondary Abs; in double staining experiments the Alexa-546-conjugated goat-anti-rabbit IgG (Invitrogen) was used.

    Techniques: Immunofluorescence, Purification, Staining, Microscopy, Software

    Co-localization of immunofluorescence signals from anti-ADAM10 pro-domain Ab and IgG from anti-ADAM10 positive patient serum Double staining was performed with rabbit anti-ADAM10 pro-domain (ab39178, Abcam), and either IgGs purified from Crc patient serum or mouse anti-HLA class I (Santa-Cruz Biotechnology). Secondary Abs were Alexa-546-conjugated goat-anti-rabbit IgG and Alexa-488-conjugated rabbit-anti-mouse IgG or FITC-conjugated goat-anti-human IgG. Cell nuclei were stained with Hoechst-33342, and differential interference contrast (DIC) images were also acquired. Staining was assessed by immunofluorescence confocal microscopy (Leica TCS SP5 Laser Scanning Confocal) and images acquired with LAS-AF (Leica) software; magnification 63X. Images from single channel and merged images are shown. Images were linearly adjusted for brightness and contrast using ImageJ 1.47v software.

    Journal: Oncotarget

    Article Title: Serological immune response against ADAM10 pro-domain is associated with favourable prognosis in stage III colorectal cancer patients

    doi: 10.18632/oncotarget.11181

    Figure Lengend Snippet: Co-localization of immunofluorescence signals from anti-ADAM10 pro-domain Ab and IgG from anti-ADAM10 positive patient serum Double staining was performed with rabbit anti-ADAM10 pro-domain (ab39178, Abcam), and either IgGs purified from Crc patient serum or mouse anti-HLA class I (Santa-Cruz Biotechnology). Secondary Abs were Alexa-546-conjugated goat-anti-rabbit IgG and Alexa-488-conjugated rabbit-anti-mouse IgG or FITC-conjugated goat-anti-human IgG. Cell nuclei were stained with Hoechst-33342, and differential interference contrast (DIC) images were also acquired. Staining was assessed by immunofluorescence confocal microscopy (Leica TCS SP5 Laser Scanning Confocal) and images acquired with LAS-AF (Leica) software; magnification 63X. Images from single channel and merged images are shown. Images were linearly adjusted for brightness and contrast using ImageJ 1.47v software.

    Article Snippet: Alexa-488-conjugated goat-anti-rabbit IgG, donkey-anti-goat IgG and rabbit-anti-mouse IgG (Invitrogen) or FITC-conjugated goat-anti-human IgG (Southern Biotechnology) were used as secondary Abs; in double staining experiments the Alexa-546-conjugated goat-anti-rabbit IgG (Invitrogen) was used.

    Techniques: Immunofluorescence, Double Staining, Purification, Staining, Confocal Microscopy, Software

    SARS-CoV RBD spike protein was internalized by the susceptible cells together with ACE2. (A) Decreased cell surface-bound RBD-Fc protein following incubation with VeroE6 cells at 37 °C compared to 4 °C. After incubation with VeroE6 cells for 3 h, cell surface-bound Fc or RBD-Fc proteins were detected using a FITC-conjugated human IgG-specific antibody. Representative FACS histograms are shown for three independent experiments. Blue and black histograms represent the results from the cells incubated with the RBD-Fc or Fc at 4 °C, respectively. The red histogram represents the results from the cells incubated with the RBD-Fc at 37 °C. (B) ACE2 internalization is the RBD spike protein-dependent. After ACE2-GFP-293 cells were incubated with the RBD-Fc or Fc protein at 37 °C for 3 h, the distribution of ACE2-GFP molecules in live cells was observed under a fluorescence microscope. (C) Co-localization of ACE2 and the RBD of SARS-CoV in ACE2-GFP-293 cells under a confocal laser-scanning microscope. After incubation with the RBD-Fc or Fc protein at 37 °C for 3 h, ACE2-GFP-293 cells were washed to remove excess RBD-Fc or Fc protein and then fixed with 4% paraformaldehyde. Immunostaining was performed using an Alexa Fluor 568-conjugated human IgG-specific antibody to detect the distribution of the Fc or RBD-Fc protein in the cells. (D) and (E) Statistical analysis of the differences between the RBD-Fc and Fc protein treatments (Chi-square test). ** P

    Journal: Virus Research

    Article Title: Endocytosis of the receptor-binding domain of SARS-CoV spike protein together with virus receptor ACE2

    doi: 10.1016/j.virusres.2008.03.004

    Figure Lengend Snippet: SARS-CoV RBD spike protein was internalized by the susceptible cells together with ACE2. (A) Decreased cell surface-bound RBD-Fc protein following incubation with VeroE6 cells at 37 °C compared to 4 °C. After incubation with VeroE6 cells for 3 h, cell surface-bound Fc or RBD-Fc proteins were detected using a FITC-conjugated human IgG-specific antibody. Representative FACS histograms are shown for three independent experiments. Blue and black histograms represent the results from the cells incubated with the RBD-Fc or Fc at 4 °C, respectively. The red histogram represents the results from the cells incubated with the RBD-Fc at 37 °C. (B) ACE2 internalization is the RBD spike protein-dependent. After ACE2-GFP-293 cells were incubated with the RBD-Fc or Fc protein at 37 °C for 3 h, the distribution of ACE2-GFP molecules in live cells was observed under a fluorescence microscope. (C) Co-localization of ACE2 and the RBD of SARS-CoV in ACE2-GFP-293 cells under a confocal laser-scanning microscope. After incubation with the RBD-Fc or Fc protein at 37 °C for 3 h, ACE2-GFP-293 cells were washed to remove excess RBD-Fc or Fc protein and then fixed with 4% paraformaldehyde. Immunostaining was performed using an Alexa Fluor 568-conjugated human IgG-specific antibody to detect the distribution of the Fc or RBD-Fc protein in the cells. (D) and (E) Statistical analysis of the differences between the RBD-Fc and Fc protein treatments (Chi-square test). ** P

    Article Snippet: Next, the cells were washed with ice-cold PBS and incubated with a FITC-conjugated affinipure goat anti-human IgG (H + L) antibody (1: 100, Jackson ImmunoResearch) at 4 °C in the dark for 30 min. After being washed with ice-cold PBS, the cells were resuspended in PBS for flow cytometry using a flow cytometer (Beckman Coulter Epics Elite ESP).

    Techniques: Incubation, FACS, Fluorescence, Microscopy, Laser-Scanning Microscopy, Immunostaining

    Colocalization of IgG and LRP2. (A–C) Immunofluorescence experiments performed on cryosections of normal human kidney tissue. (A) Staining for LRP2 (red) using a rabbit polyclonal antibody along the apical membrane of the tubular epithelium. (B) Indirect immunofluorescence of serum from a patient with ABBA on the normal human kidney. (C) Strong colocalization of LRP2 and serum antibodies from a patient with ABBA. (D–F) Immunofluorescence experiments performed on a renal biopsy sample from a patient with anti-LRP2 nephropathy shows granular TBM staining in a proximal tubule for LRP2 using (D) a mouse mAb and (E) IgG. (F) There is strong colocalization of LRP2 and IgG in the TBM deposits. (G–I) Immunofluorescence experiments performed on a renal biopsy sample from a patient with lupus nephritis shows staining of the tubular apical membrane for LRP2 using (G) a mouse mAb and (H) granular TBM staining for IgG. (I) There is complete lack of colocalization in a patient without anti-LRP2 nephropathy.

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: LDL Receptor-Related Protein 2 (Megalin) as a Target Antigen in Human Kidney Anti-Brush Border Antibody Disease

    doi: 10.1681/ASN.2017060664

    Figure Lengend Snippet: Colocalization of IgG and LRP2. (A–C) Immunofluorescence experiments performed on cryosections of normal human kidney tissue. (A) Staining for LRP2 (red) using a rabbit polyclonal antibody along the apical membrane of the tubular epithelium. (B) Indirect immunofluorescence of serum from a patient with ABBA on the normal human kidney. (C) Strong colocalization of LRP2 and serum antibodies from a patient with ABBA. (D–F) Immunofluorescence experiments performed on a renal biopsy sample from a patient with anti-LRP2 nephropathy shows granular TBM staining in a proximal tubule for LRP2 using (D) a mouse mAb and (E) IgG. (F) There is strong colocalization of LRP2 and IgG in the TBM deposits. (G–I) Immunofluorescence experiments performed on a renal biopsy sample from a patient with lupus nephritis shows staining of the tubular apical membrane for LRP2 using (G) a mouse mAb and (H) granular TBM staining for IgG. (I) There is complete lack of colocalization in a patient without anti-LRP2 nephropathy.

    Article Snippet: The sections were reacted with mouse monoclonal anti-human LRP2 (1:1000; EMD Millipore, Billerica, MA), then polyclonal (Rhodamine Red-X) goat anti-mouse IgG Fc γ subclass 1 (Jackson ImmunoResearch, West Grove, PA), and polyclonal (FITC-conjugated) goat anti-human IgG (1:100; Kent Laboratories, Bellingham, WA).

    Techniques: Immunofluorescence, Staining