phycoerythrin pe conjugated anti mouse immunoglobulin g  (Jackson Immuno)

 
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    Name:
    R Phycoerythrin AffiniPure F ab ₂ Fragment Goat Anti Mouse IgG Fcγ fragment specific
    Description:
    F ab 2 fragment antibodies are generated by pepsin digestion of whole IgG antibodies to remove most of the Fc region while leaving some of the hinge region F ab 2 fragments have two antigen binding Fab portions linked together by disulfide bonds and therefore they are divalent The average molecular weight is about 110 kDa They are used for specific applications such as to avoid binding of secondary antibodies to live cells with Fc receptors or to Protein A or Protein G Based on immunoelectrophoresis and or ELISA the antibody reacts with the Fc portion of mouse IgG heavy chain but not with the Fab portion of mouse immunoglobulins No antibody was detected against mouse IgM or non immunoglobulin serum proteins The antibody has been tested by ELISA and or solid phase adsorbed to ensure minimal cross reaction with human bovine and horse serum proteins but it may cross react with immunoglobulins from other species
    Catalog Number:
    115-116-071
    Price:
    249.0
    Category:
    Fluorescent Protein Conjugates R PE APC and PerCP
    Conjugate:
    R Phycoerythrin
    Size:
    1 0 ml
    Format:
    F(ab')₂ Fragment
    Host:
    Goat
    Buy from Supplier


    Structured Review

    Jackson Immuno phycoerythrin pe conjugated anti mouse immunoglobulin g
    Antibody-mediated inhibition of cellular HCV-LP binding. HCV-LPs were incubated with anti-HCV-positive serum or a pool of anti-HCV-negative control sera (dilution, 1:50). HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. After removal of nonbound HCV-LP-antibody complexes by washing the cells in PBS-2% BSA, the binding of HCV-LPs was detected by flow cytometry using the mouse monoclonal anti-E2 antibody AP33 and PE-conjugated anti-mouse <t>IgG</t> (A and B) or human polyclonal anti-HCV and FITC-conjugated anti-human IgG antibody (C). The fluorescence intensity and relative cell numbers (Counts) are shown on the x and y axes, respectively. NC, negative control, corresponding to HuH-7 cells incubated with control insect cell preparations (GUS) and control serum. (D) For the assessment of concentration-dependent antibody-mediated inhibition of binding, HCV-LPs (genotype 1b) were incubated in subsaturating concentrations with an anti-HCV-positive serum or a pool of anti-HCV-negative control sera (at various dilutions as indicated on the x axis) for 1 h at 37°C. HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. HCV-LP binding to the HuH-7 cells was detected by flow cytometry as described above. Inhibition of cellular HCV-LP binding ( y axis) was calculated as described in Materials and Methods.
    F ab 2 fragment antibodies are generated by pepsin digestion of whole IgG antibodies to remove most of the Fc region while leaving some of the hinge region F ab 2 fragments have two antigen binding Fab portions linked together by disulfide bonds and therefore they are divalent The average molecular weight is about 110 kDa They are used for specific applications such as to avoid binding of secondary antibodies to live cells with Fc receptors or to Protein A or Protein G Based on immunoelectrophoresis and or ELISA the antibody reacts with the Fc portion of mouse IgG heavy chain but not with the Fab portion of mouse immunoglobulins No antibody was detected against mouse IgM or non immunoglobulin serum proteins The antibody has been tested by ELISA and or solid phase adsorbed to ensure minimal cross reaction with human bovine and horse serum proteins but it may cross react with immunoglobulins from other species
    https://www.bioz.com/result/phycoerythrin pe conjugated anti mouse immunoglobulin g/product/Jackson Immuno
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phycoerythrin pe conjugated anti mouse immunoglobulin g - by Bioz Stars, 2021-06
    94/100 stars

    Images

    1) Product Images from "Inhibition of Hepatitis C Virus-Like Particle Binding to Target Cells by Antiviral Antibodies in Acute and Chronic Hepatitis C"

    Article Title: Inhibition of Hepatitis C Virus-Like Particle Binding to Target Cells by Antiviral Antibodies in Acute and Chronic Hepatitis C

    Journal: Journal of Virology

    doi: 10.1128/JVI.78.17.9030-9040.2004

    Antibody-mediated inhibition of cellular HCV-LP binding. HCV-LPs were incubated with anti-HCV-positive serum or a pool of anti-HCV-negative control sera (dilution, 1:50). HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. After removal of nonbound HCV-LP-antibody complexes by washing the cells in PBS-2% BSA, the binding of HCV-LPs was detected by flow cytometry using the mouse monoclonal anti-E2 antibody AP33 and PE-conjugated anti-mouse IgG (A and B) or human polyclonal anti-HCV and FITC-conjugated anti-human IgG antibody (C). The fluorescence intensity and relative cell numbers (Counts) are shown on the x and y axes, respectively. NC, negative control, corresponding to HuH-7 cells incubated with control insect cell preparations (GUS) and control serum. (D) For the assessment of concentration-dependent antibody-mediated inhibition of binding, HCV-LPs (genotype 1b) were incubated in subsaturating concentrations with an anti-HCV-positive serum or a pool of anti-HCV-negative control sera (at various dilutions as indicated on the x axis) for 1 h at 37°C. HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. HCV-LP binding to the HuH-7 cells was detected by flow cytometry as described above. Inhibition of cellular HCV-LP binding ( y axis) was calculated as described in Materials and Methods.
    Figure Legend Snippet: Antibody-mediated inhibition of cellular HCV-LP binding. HCV-LPs were incubated with anti-HCV-positive serum or a pool of anti-HCV-negative control sera (dilution, 1:50). HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. After removal of nonbound HCV-LP-antibody complexes by washing the cells in PBS-2% BSA, the binding of HCV-LPs was detected by flow cytometry using the mouse monoclonal anti-E2 antibody AP33 and PE-conjugated anti-mouse IgG (A and B) or human polyclonal anti-HCV and FITC-conjugated anti-human IgG antibody (C). The fluorescence intensity and relative cell numbers (Counts) are shown on the x and y axes, respectively. NC, negative control, corresponding to HuH-7 cells incubated with control insect cell preparations (GUS) and control serum. (D) For the assessment of concentration-dependent antibody-mediated inhibition of binding, HCV-LPs (genotype 1b) were incubated in subsaturating concentrations with an anti-HCV-positive serum or a pool of anti-HCV-negative control sera (at various dilutions as indicated on the x axis) for 1 h at 37°C. HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. HCV-LP binding to the HuH-7 cells was detected by flow cytometry as described above. Inhibition of cellular HCV-LP binding ( y axis) was calculated as described in Materials and Methods.

    Techniques Used: Inhibition, Binding Assay, Incubation, Negative Control, Flow Cytometry, Cytometry, Fluorescence, Concentration Assay

    Related Articles

    Staining:

    Article Title: The Streptococcus agalactiae complement interfering protein combines multiple complement-inhibitory mechanisms by interacting with both C4 and C3 ligands
    Article Snippet: The bC3d–SA or CIP–bC3d–SA or Fib–bC3d–SA complexes were then added to Raji cells or B cells purified from human PBMCs (5 × 105/sample), and the mixture was left 15 min at 37°C and washed with PBS. .. Bound C3d was then stained with a mouse anti-C3d mAb (prediluted 1:100; Thermo Fisher Scientific) and revealed with a phycoerythrin-labeled anti-mouse secondary antibody (Jackson ImmunoResearch Laboratories, West Grove, PA, USA). .. Both incubations were conducted for 10 min at 37°C, and after each incubation, 2 washes in PBS–fetal calf serum 1% (v/v) were performed.

    Article Title: Separation and Characterization of Epithelial and Mesenchymal-like Murine Mammary Tumor Cells Reveals Epithelial Cell Differentiation Plasticity and Enhanced Tumorigenicity of Epithelial-enriched Tumor Cells
    Article Snippet: .. To obtain purified cultures of Neu+ mammary tumor cells, cells were stained with anti-c-ErbB2/c-neu mouse monoclonal antibody clone 7.16.4 (Calbiochem-EMD Chemicals, Gibbstown, NJ) and phycoerythrin (PE)-conjugated goat anti-mouse (Jackson ImmunoResearch, West Grove, PA) secondary antibody, and incubated with anti-PE microbeads (Milteni Biotech, Auburn, CA). .. Cells were then separated by positive selection using automated immunomagnetic sorting (AutoMACS, Milteni Biotech).

    Article Title: p21-activated kinase 4 interacts with integrin ?v?5 and regulates ?v?5-mediated cell migration
    Article Snippet: MCF-7 cells stably transfected with EGFP–PAK4 or EGFP control were plated into the wells in triplicate at 5 ×104 cells/well in cell adhesion buffer (RPMI 1640, 2 mM CaCl2 , 1 mM MgCl2 , 0.2 mM MnCl2, and 0.5% BSA) and allowed to attach for 60 min. After careful washing of nonbound cells using adhesion buffer, MTT was used to quantify the number of stably transfected cells attached. .. Flow cytometry analyses The efficiency for cell transfections and the cell surface expression levels of integrins were analyzed by measurement of EGFP content and phycoerythrin staining intensity, respectively, by FACScan® flow cytometer using CellQuest software (Becton Dickinson) after staining with anti–integrin αvβ5 mAb P1F6 and a phycoerythrin-conjugated secondary goat anti–mouse pAb (Jackson ImmunoResearch Laboratories). ..

    Purification:

    Article Title: Separation and Characterization of Epithelial and Mesenchymal-like Murine Mammary Tumor Cells Reveals Epithelial Cell Differentiation Plasticity and Enhanced Tumorigenicity of Epithelial-enriched Tumor Cells
    Article Snippet: .. To obtain purified cultures of Neu+ mammary tumor cells, cells were stained with anti-c-ErbB2/c-neu mouse monoclonal antibody clone 7.16.4 (Calbiochem-EMD Chemicals, Gibbstown, NJ) and phycoerythrin (PE)-conjugated goat anti-mouse (Jackson ImmunoResearch, West Grove, PA) secondary antibody, and incubated with anti-PE microbeads (Milteni Biotech, Auburn, CA). .. Cells were then separated by positive selection using automated immunomagnetic sorting (AutoMACS, Milteni Biotech).

    Incubation:

    Article Title: Separation and Characterization of Epithelial and Mesenchymal-like Murine Mammary Tumor Cells Reveals Epithelial Cell Differentiation Plasticity and Enhanced Tumorigenicity of Epithelial-enriched Tumor Cells
    Article Snippet: .. To obtain purified cultures of Neu+ mammary tumor cells, cells were stained with anti-c-ErbB2/c-neu mouse monoclonal antibody clone 7.16.4 (Calbiochem-EMD Chemicals, Gibbstown, NJ) and phycoerythrin (PE)-conjugated goat anti-mouse (Jackson ImmunoResearch, West Grove, PA) secondary antibody, and incubated with anti-PE microbeads (Milteni Biotech, Auburn, CA). .. Cells were then separated by positive selection using automated immunomagnetic sorting (AutoMACS, Milteni Biotech).

    Article Title: Development and Evaluation of a Broad Bead-Based Multiplex Immunoassay To Measure IgG Seroreactivity against Human Polyomaviruses
    Article Snippet: .. The coupling of the complete GST-VP1.tag fusion protein to the bead was verified on a Bio-Plex apparatus using mouse anti-tag antibodies (1:100; a kind gift from M. Pawlita), followed by anti-mouse immunoglobulin-phycoerythrin for detection (1:250; Jackson ImmunoResearch Laboratories Inc., West Grove, PA, USA) (which were incubated for 30 min each in the dark at room temperature). .. In the HPyV multiplex immunoassay, serum samples (1:100) were incubated for 1 h in blocking buffer (1 mg/ml casein, 0.5% polyvinyl alcohol, 0.8% polyvinylpyrrolidone, 2.5% Super ChemiBlock [Chemicon International, Billerica, MA, USA], 2 mg/ml GST bacterial lysate in phosphate-buffered saline) to suppress potential nonspecific binding to the beads or to GST ( , ).

    Article Title: Inhibition of Hepatitis C Virus-Like Particle Binding to Target Cells by Antiviral Antibodies in Acute and Chronic Hepatitis C
    Article Snippet: The cells were washed three times in PBS-2% BSA at 4°C. .. The cells were incubated for 30 min at 4°C with phycoerythrin (PE)-conjugated anti-mouse immunoglobulin G (IgG) antibody (Jackson ImmunoResearch, West Grove, Pa.) or fluorescein isothiocyanate (FITC)-conjugated anti-human IgG antibody (ICN Biomedicals Inc., Aurora, Ohio) diluted in 50 μl of PBS-2% BSA (1:100). .. The cells were washed three times in PBS at 4°C.

    Flow Cytometry:

    Article Title: p21-activated kinase 4 interacts with integrin ?v?5 and regulates ?v?5-mediated cell migration
    Article Snippet: MCF-7 cells stably transfected with EGFP–PAK4 or EGFP control were plated into the wells in triplicate at 5 ×104 cells/well in cell adhesion buffer (RPMI 1640, 2 mM CaCl2 , 1 mM MgCl2 , 0.2 mM MnCl2, and 0.5% BSA) and allowed to attach for 60 min. After careful washing of nonbound cells using adhesion buffer, MTT was used to quantify the number of stably transfected cells attached. .. Flow cytometry analyses The efficiency for cell transfections and the cell surface expression levels of integrins were analyzed by measurement of EGFP content and phycoerythrin staining intensity, respectively, by FACScan® flow cytometer using CellQuest software (Becton Dickinson) after staining with anti–integrin αvβ5 mAb P1F6 and a phycoerythrin-conjugated secondary goat anti–mouse pAb (Jackson ImmunoResearch Laboratories). ..

    Transfection:

    Article Title: p21-activated kinase 4 interacts with integrin ?v?5 and regulates ?v?5-mediated cell migration
    Article Snippet: MCF-7 cells stably transfected with EGFP–PAK4 or EGFP control were plated into the wells in triplicate at 5 ×104 cells/well in cell adhesion buffer (RPMI 1640, 2 mM CaCl2 , 1 mM MgCl2 , 0.2 mM MnCl2, and 0.5% BSA) and allowed to attach for 60 min. After careful washing of nonbound cells using adhesion buffer, MTT was used to quantify the number of stably transfected cells attached. .. Flow cytometry analyses The efficiency for cell transfections and the cell surface expression levels of integrins were analyzed by measurement of EGFP content and phycoerythrin staining intensity, respectively, by FACScan® flow cytometer using CellQuest software (Becton Dickinson) after staining with anti–integrin αvβ5 mAb P1F6 and a phycoerythrin-conjugated secondary goat anti–mouse pAb (Jackson ImmunoResearch Laboratories). ..

    Expressing:

    Article Title: p21-activated kinase 4 interacts with integrin ?v?5 and regulates ?v?5-mediated cell migration
    Article Snippet: MCF-7 cells stably transfected with EGFP–PAK4 or EGFP control were plated into the wells in triplicate at 5 ×104 cells/well in cell adhesion buffer (RPMI 1640, 2 mM CaCl2 , 1 mM MgCl2 , 0.2 mM MnCl2, and 0.5% BSA) and allowed to attach for 60 min. After careful washing of nonbound cells using adhesion buffer, MTT was used to quantify the number of stably transfected cells attached. .. Flow cytometry analyses The efficiency for cell transfections and the cell surface expression levels of integrins were analyzed by measurement of EGFP content and phycoerythrin staining intensity, respectively, by FACScan® flow cytometer using CellQuest software (Becton Dickinson) after staining with anti–integrin αvβ5 mAb P1F6 and a phycoerythrin-conjugated secondary goat anti–mouse pAb (Jackson ImmunoResearch Laboratories). ..

    Software:

    Article Title: p21-activated kinase 4 interacts with integrin ?v?5 and regulates ?v?5-mediated cell migration
    Article Snippet: MCF-7 cells stably transfected with EGFP–PAK4 or EGFP control were plated into the wells in triplicate at 5 ×104 cells/well in cell adhesion buffer (RPMI 1640, 2 mM CaCl2 , 1 mM MgCl2 , 0.2 mM MnCl2, and 0.5% BSA) and allowed to attach for 60 min. After careful washing of nonbound cells using adhesion buffer, MTT was used to quantify the number of stably transfected cells attached. .. Flow cytometry analyses The efficiency for cell transfections and the cell surface expression levels of integrins were analyzed by measurement of EGFP content and phycoerythrin staining intensity, respectively, by FACScan® flow cytometer using CellQuest software (Becton Dickinson) after staining with anti–integrin αvβ5 mAb P1F6 and a phycoerythrin-conjugated secondary goat anti–mouse pAb (Jackson ImmunoResearch Laboratories). ..

    FACS:

    Article Title: Adenovirus Serotype 30 Fiber Does Not Mediate Transduction via the Coxsackie-Adenovirus Receptor
    Article Snippet: To detect CAR expression, cells were detached from dishes with EDTA, spun down, resuspended in 1% fetal bovine serum-PBS at 2 × 105 cells/ml, and incubated with monoclonal antibodies against CAR (RmcB; a kind gift from Jeffrey Bergelson, Children’s Hospital of Philadelphia, Philadelphia, Pa.) ( , ) for 45 min at 37°C. .. Cells were pelleted, washed, and resuspended with R -phycoerythrin-conjugated goat anti-mouse secondary antibodies (Jackson ImmunoResearch) for 45 min at 4°C, prior to FACS analysis. .. CHO cells were transfected with AdhCAR previously prepared with CaPi as described above.

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  • 94
    Jackson Immuno phycoerythrin pe conjugated anti mouse immunoglobulin g
    Antibody-mediated inhibition of cellular HCV-LP binding. HCV-LPs were incubated with anti-HCV-positive serum or a pool of anti-HCV-negative control sera (dilution, 1:50). HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. After removal of nonbound HCV-LP-antibody complexes by washing the cells in PBS-2% BSA, the binding of HCV-LPs was detected by flow cytometry using the mouse monoclonal anti-E2 antibody AP33 and PE-conjugated anti-mouse <t>IgG</t> (A and B) or human polyclonal anti-HCV and FITC-conjugated anti-human IgG antibody (C). The fluorescence intensity and relative cell numbers (Counts) are shown on the x and y axes, respectively. NC, negative control, corresponding to HuH-7 cells incubated with control insect cell preparations (GUS) and control serum. (D) For the assessment of concentration-dependent antibody-mediated inhibition of binding, HCV-LPs (genotype 1b) were incubated in subsaturating concentrations with an anti-HCV-positive serum or a pool of anti-HCV-negative control sera (at various dilutions as indicated on the x axis) for 1 h at 37°C. HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. HCV-LP binding to the HuH-7 cells was detected by flow cytometry as described above. Inhibition of cellular HCV-LP binding ( y axis) was calculated as described in Materials and Methods.
    Phycoerythrin Pe Conjugated Anti Mouse Immunoglobulin G, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phycoerythrin pe conjugated anti mouse immunoglobulin g/product/Jackson Immuno
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phycoerythrin pe conjugated anti mouse immunoglobulin g - by Bioz Stars, 2021-06
    94/100 stars
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    94
    Jackson Immuno pe conjugated goat anti mouse immunoglobulin g igg
    Expression of CXCR4 in CD4 + , CD4 + CD25 + , and CD4 + CD25 − cells. (A) LN-derived cells (5 × 10 5 ) were incubated with cross-reacting anti-CXCR4 antibody (44717) followed by PE-conjugated goat anti-mouse <t>IgG.</t> The cells were then washed, stained with FITC-labeled feline anti-CD4 antibody (30A), and analyzed by flow cytometry. Numbers represent the percentages of cells within each quadrant. Ab, antibody. (B) LN-derived cells were sorted into CD4 + CD25 + and CD4 + CD25 − populations. Purified cells (3 × 10 5 ) were stained for surface CXCR4 expression as described above either immediately postisolation or after stimulation for 3 days with 100-U/ml IL-2 or 2-μg/ml ConA and analyzed by flow cytometry. One representative of four independent experiments is shown. Shaded region, secondary antibody (Ab) alone.
    Pe Conjugated Goat Anti Mouse Immunoglobulin G Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pe conjugated goat anti mouse immunoglobulin g igg/product/Jackson Immuno
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pe conjugated goat anti mouse immunoglobulin g igg - by Bioz Stars, 2021-06
    94/100 stars
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    93
    Jackson Immuno r phycoerythrin pe conjugated donkey f ab 2 anti mouse immunoglobulin g igg
    Expression of CXCR4 in CD4 + , CD4 + CD25 + , and CD4 + CD25 − cells. (A) LN-derived cells (5 × 10 5 ) were incubated with cross-reacting anti-CXCR4 antibody (44717) followed by PE-conjugated goat anti-mouse <t>IgG.</t> The cells were then washed, stained with FITC-labeled feline anti-CD4 antibody (30A), and analyzed by flow cytometry. Numbers represent the percentages of cells within each quadrant. Ab, antibody. (B) LN-derived cells were sorted into CD4 + CD25 + and CD4 + CD25 − populations. Purified cells (3 × 10 5 ) were stained for surface CXCR4 expression as described above either immediately postisolation or after stimulation for 3 days with 100-U/ml IL-2 or 2-μg/ml ConA and analyzed by flow cytometry. One representative of four independent experiments is shown. Shaded region, secondary antibody (Ab) alone.
    R Phycoerythrin Pe Conjugated Donkey F Ab 2 Anti Mouse Immunoglobulin G Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/r phycoerythrin pe conjugated donkey f ab 2 anti mouse immunoglobulin g igg/product/Jackson Immuno
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    r phycoerythrin pe conjugated donkey f ab 2 anti mouse immunoglobulin g igg - by Bioz Stars, 2021-06
    93/100 stars
      Buy from Supplier

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    Antibody-mediated inhibition of cellular HCV-LP binding. HCV-LPs were incubated with anti-HCV-positive serum or a pool of anti-HCV-negative control sera (dilution, 1:50). HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. After removal of nonbound HCV-LP-antibody complexes by washing the cells in PBS-2% BSA, the binding of HCV-LPs was detected by flow cytometry using the mouse monoclonal anti-E2 antibody AP33 and PE-conjugated anti-mouse IgG (A and B) or human polyclonal anti-HCV and FITC-conjugated anti-human IgG antibody (C). The fluorescence intensity and relative cell numbers (Counts) are shown on the x and y axes, respectively. NC, negative control, corresponding to HuH-7 cells incubated with control insect cell preparations (GUS) and control serum. (D) For the assessment of concentration-dependent antibody-mediated inhibition of binding, HCV-LPs (genotype 1b) were incubated in subsaturating concentrations with an anti-HCV-positive serum or a pool of anti-HCV-negative control sera (at various dilutions as indicated on the x axis) for 1 h at 37°C. HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. HCV-LP binding to the HuH-7 cells was detected by flow cytometry as described above. Inhibition of cellular HCV-LP binding ( y axis) was calculated as described in Materials and Methods.

    Journal: Journal of Virology

    Article Title: Inhibition of Hepatitis C Virus-Like Particle Binding to Target Cells by Antiviral Antibodies in Acute and Chronic Hepatitis C

    doi: 10.1128/JVI.78.17.9030-9040.2004

    Figure Lengend Snippet: Antibody-mediated inhibition of cellular HCV-LP binding. HCV-LPs were incubated with anti-HCV-positive serum or a pool of anti-HCV-negative control sera (dilution, 1:50). HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. After removal of nonbound HCV-LP-antibody complexes by washing the cells in PBS-2% BSA, the binding of HCV-LPs was detected by flow cytometry using the mouse monoclonal anti-E2 antibody AP33 and PE-conjugated anti-mouse IgG (A and B) or human polyclonal anti-HCV and FITC-conjugated anti-human IgG antibody (C). The fluorescence intensity and relative cell numbers (Counts) are shown on the x and y axes, respectively. NC, negative control, corresponding to HuH-7 cells incubated with control insect cell preparations (GUS) and control serum. (D) For the assessment of concentration-dependent antibody-mediated inhibition of binding, HCV-LPs (genotype 1b) were incubated in subsaturating concentrations with an anti-HCV-positive serum or a pool of anti-HCV-negative control sera (at various dilutions as indicated on the x axis) for 1 h at 37°C. HCV-LP-antibody complexes were added to HuH-7 cells for 1 h at 4°C. HCV-LP binding to the HuH-7 cells was detected by flow cytometry as described above. Inhibition of cellular HCV-LP binding ( y axis) was calculated as described in Materials and Methods.

    Article Snippet: The cells were incubated for 30 min at 4°C with phycoerythrin (PE)-conjugated anti-mouse immunoglobulin G (IgG) antibody (Jackson ImmunoResearch, West Grove, Pa.) or fluorescein isothiocyanate (FITC)-conjugated anti-human IgG antibody (ICN Biomedicals Inc., Aurora, Ohio) diluted in 50 μl of PBS-2% BSA (1:100).

    Techniques: Inhibition, Binding Assay, Incubation, Negative Control, Flow Cytometry, Cytometry, Fluorescence, Concentration Assay

    Expression of CXCR4 in CD4 + , CD4 + CD25 + , and CD4 + CD25 − cells. (A) LN-derived cells (5 × 10 5 ) were incubated with cross-reacting anti-CXCR4 antibody (44717) followed by PE-conjugated goat anti-mouse IgG. The cells were then washed, stained with FITC-labeled feline anti-CD4 antibody (30A), and analyzed by flow cytometry. Numbers represent the percentages of cells within each quadrant. Ab, antibody. (B) LN-derived cells were sorted into CD4 + CD25 + and CD4 + CD25 − populations. Purified cells (3 × 10 5 ) were stained for surface CXCR4 expression as described above either immediately postisolation or after stimulation for 3 days with 100-U/ml IL-2 or 2-μg/ml ConA and analyzed by flow cytometry. One representative of four independent experiments is shown. Shaded region, secondary antibody (Ab) alone.

    Journal: Journal of Virology

    Article Title: Preferential Feline Immunodeficiency Virus (FIV) Infection of CD4+ CD25+ T-Regulatory Cells Correlates both with Surface Expression of CXCR4 and Activation of FIV Long Terminal Repeat Binding Cellular Transcriptional Factors

    doi: 10.1128/JVI.79.8.4965-4976.2005

    Figure Lengend Snippet: Expression of CXCR4 in CD4 + , CD4 + CD25 + , and CD4 + CD25 − cells. (A) LN-derived cells (5 × 10 5 ) were incubated with cross-reacting anti-CXCR4 antibody (44717) followed by PE-conjugated goat anti-mouse IgG. The cells were then washed, stained with FITC-labeled feline anti-CD4 antibody (30A), and analyzed by flow cytometry. Numbers represent the percentages of cells within each quadrant. Ab, antibody. (B) LN-derived cells were sorted into CD4 + CD25 + and CD4 + CD25 − populations. Purified cells (3 × 10 5 ) were stained for surface CXCR4 expression as described above either immediately postisolation or after stimulation for 3 days with 100-U/ml IL-2 or 2-μg/ml ConA and analyzed by flow cytometry. One representative of four independent experiments is shown. Shaded region, secondary antibody (Ab) alone.

    Article Snippet: Cell surface CXCR4 expression was determined using the cross-reacting human anti-CXCR4 antibody (44717) followed by PE-conjugated goat anti-mouse immunoglobulin G (IgG) (Jackson ImmunoResearch Laboratories, West Grove, Pa.).

    Techniques: Expressing, Derivative Assay, Incubation, Staining, Labeling, Flow Cytometry, Cytometry, Purification