anti h7n9 hemagglutinin ha antibody rabbit pab  (Sino Biological)


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    Name:
    Anti H7N9 Hemagglutinin HA Antibody Rabbit PAb
    Description:
    Produced in rabbits immunized with purified recombinant Influenza A H7N9 A Anhui 1 2013 HA Catalog 40103 V08B EPI439507 Met1 Val524 HA specific IgG was purified by Influenza A H7N9 A Anhui 1 2013 HA affinity chromatography
    Catalog Number:
    40103-RP02
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    H7N9
    Applications:
    WB,ELISA
    Immunogen:
    Recombinant Influenza A H7N9 (A/Anhui/1/2013) HA protein (Catalog#40103-V08B)
    Antibody Type:
    PAb
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Sino Biological anti h7n9 hemagglutinin ha antibody rabbit pab
    Biological and chemical modifications of <t>H7N9</t> VLPs. (A) SDS-PAGE and western blot analysis of biologically modified H7N9 VLPs. Lane 1–3 represent 320 ng, 180 ng and 90 ng of αGal (-) VLP based on HA content, respectively. Lane 4–6 are 130 ng, 90, and 40 ng of αGal (+) VLP, respectively. (B) SDS-PAGE and western blot analysis of chemically modified H7N9 VLPs. S: protein standards; 1: αGal (-) H7N9 VLP; 2: intermediate of αGal (-) H7N9 VLP reacted with GO; 3: H7N9 conjugated with αGal linker a11; 4: H7N9 conjugated with αGal linker aN11; 5: H7N9 VLPs conjugated with control linker sp11. (C) Electron microscopy of H7N9 VLPs. (D) Electron microscopy of H7N9 VLPs stained with gold particles. Negative: VLPs were only stained with immunogold donkey anti-rabbit secondary antibody; Positive: VLPs were stained with anti-HA7 rabbit polyclonal antibody, followed by staining with immunogold donkey anti-rabbit secondary antibody. Sp11 is the aminooxy spacer control linker without αGal antigen; a11 is one type of αGal (Galα1,3-Galβ1,4-Glc) aminooxy linker; aN11 is the other type of αGal (Galα1,3-Galβ1,4-GlcNAc) aminooxy linker.
    Produced in rabbits immunized with purified recombinant Influenza A H7N9 A Anhui 1 2013 HA Catalog 40103 V08B EPI439507 Met1 Val524 HA specific IgG was purified by Influenza A H7N9 A Anhui 1 2013 HA affinity chromatography
    https://www.bioz.com/result/anti h7n9 hemagglutinin ha antibody rabbit pab/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti h7n9 hemagglutinin ha antibody rabbit pab - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Addition of αGal HyperAcute™ technology to recombinant avian influenza vaccines induces strong low-dose antibody responses"

    Article Title: Addition of αGal HyperAcute™ technology to recombinant avian influenza vaccines induces strong low-dose antibody responses

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0182683

    Biological and chemical modifications of H7N9 VLPs. (A) SDS-PAGE and western blot analysis of biologically modified H7N9 VLPs. Lane 1–3 represent 320 ng, 180 ng and 90 ng of αGal (-) VLP based on HA content, respectively. Lane 4–6 are 130 ng, 90, and 40 ng of αGal (+) VLP, respectively. (B) SDS-PAGE and western blot analysis of chemically modified H7N9 VLPs. S: protein standards; 1: αGal (-) H7N9 VLP; 2: intermediate of αGal (-) H7N9 VLP reacted with GO; 3: H7N9 conjugated with αGal linker a11; 4: H7N9 conjugated with αGal linker aN11; 5: H7N9 VLPs conjugated with control linker sp11. (C) Electron microscopy of H7N9 VLPs. (D) Electron microscopy of H7N9 VLPs stained with gold particles. Negative: VLPs were only stained with immunogold donkey anti-rabbit secondary antibody; Positive: VLPs were stained with anti-HA7 rabbit polyclonal antibody, followed by staining with immunogold donkey anti-rabbit secondary antibody. Sp11 is the aminooxy spacer control linker without αGal antigen; a11 is one type of αGal (Galα1,3-Galβ1,4-Glc) aminooxy linker; aN11 is the other type of αGal (Galα1,3-Galβ1,4-GlcNAc) aminooxy linker.
    Figure Legend Snippet: Biological and chemical modifications of H7N9 VLPs. (A) SDS-PAGE and western blot analysis of biologically modified H7N9 VLPs. Lane 1–3 represent 320 ng, 180 ng and 90 ng of αGal (-) VLP based on HA content, respectively. Lane 4–6 are 130 ng, 90, and 40 ng of αGal (+) VLP, respectively. (B) SDS-PAGE and western blot analysis of chemically modified H7N9 VLPs. S: protein standards; 1: αGal (-) H7N9 VLP; 2: intermediate of αGal (-) H7N9 VLP reacted with GO; 3: H7N9 conjugated with αGal linker a11; 4: H7N9 conjugated with αGal linker aN11; 5: H7N9 VLPs conjugated with control linker sp11. (C) Electron microscopy of H7N9 VLPs. (D) Electron microscopy of H7N9 VLPs stained with gold particles. Negative: VLPs were only stained with immunogold donkey anti-rabbit secondary antibody; Positive: VLPs were stained with anti-HA7 rabbit polyclonal antibody, followed by staining with immunogold donkey anti-rabbit secondary antibody. Sp11 is the aminooxy spacer control linker without αGal antigen; a11 is one type of αGal (Galα1,3-Galβ1,4-Glc) aminooxy linker; aN11 is the other type of αGal (Galα1,3-Galβ1,4-GlcNAc) aminooxy linker.

    Techniques Used: SDS Page, Western Blot, Modification, Electron Microscopy, Staining

    Related Articles

    Recombinant:

    Article Title: Rapid, Sensitive, and Selective Detection of H5 Hemagglutinin from Avian Influenza Virus Using an Immunowall Device
    Article Snippet: .. Mouse anti-H5N1 HA monoclonal antibody, recombinant H5N1 HA, recombinant H1N1 HA, recombinant H3N2 HA, recombinant H7N9 HA, and rabbit anti-avian influenza A HA polyclonal antibody were purchased from Sino Biological, Inc. (Beijing, China). ..

    Article Title: Production of influenza virus-like particles using recombinant insect cells
    Article Snippet: .. 2.7 ImmunoprecipitationCulture supernatant of recombinant cells was mixed with either rabbit anti-HA polyclonal antibody (Sino Biological) or rabbit anti-M1 polyclonal antibody (Sino Biological) in microtubes that were then incubated for 1 h. After protein G PLUS-agarose beads (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were added, the microtubes were rotated for 1 h. Bead-antigen complexes were then pelleted by centrifugation, and the pellet was washed 3 times with PBS. ..

    Incubation:

    Article Title: Production of influenza virus-like particles using recombinant insect cells
    Article Snippet: .. 2.7 ImmunoprecipitationCulture supernatant of recombinant cells was mixed with either rabbit anti-HA polyclonal antibody (Sino Biological) or rabbit anti-M1 polyclonal antibody (Sino Biological) in microtubes that were then incubated for 1 h. After protein G PLUS-agarose beads (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were added, the microtubes were rotated for 1 h. Bead-antigen complexes were then pelleted by centrifugation, and the pellet was washed 3 times with PBS. ..

    Centrifugation:

    Article Title: Production of influenza virus-like particles using recombinant insect cells
    Article Snippet: .. 2.7 ImmunoprecipitationCulture supernatant of recombinant cells was mixed with either rabbit anti-HA polyclonal antibody (Sino Biological) or rabbit anti-M1 polyclonal antibody (Sino Biological) in microtubes that were then incubated for 1 h. After protein G PLUS-agarose beads (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were added, the microtubes were rotated for 1 h. Bead-antigen complexes were then pelleted by centrifugation, and the pellet was washed 3 times with PBS. ..

    FLAG-tag:

    Article Title: Transcriptome profiling and protease inhibition experiments identify proteases that activate H3N2 influenza A and influenza B viruses in murine airways
    Article Snippet: .. Antibodies used in this study were a polyclonal rabbit antibody against HA1 of A/TW/3446/02 (H3N2) (GeneTex, GTX127363), a polyclonal rabbit antibody against HA1 of B/Brisbane/60/2008 (Sino Biological, 40016-T38), a polyclonal rabbit antibody against HA of A/Anhui/1/13 (H7N9) (Sino Biological, 40103RP02), a polyclonal rabbit antibody against HA of H9N2 (Thermo Fisher Scientific, PA5–81658), a monoclonal mouse antibody against β-actin (Abcam, ab6276), a monoclonal mouse antibody against IBV NP (Thermo Fisher Scientific, B017 (B35G)), a monoclonal mouse antibody against IAV NP (Abcam, ab43821), a polyclonal rabbit antibody against FLAG tag (Sigma–Aldrich), and a polyclonal goat antibody against mouse tryptase ϵ (Thermo Fisher Scientific, PA5–47245). ..

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    Sino Biological rabbit anti ha polyclonal antibody
    Western blot analysis of culture supernatant immunoprecipitated with rabbit anti-HA <t>polyclonal</t> antibody. Mouse anti-HA monoclonal antibody (a) and mouse anti-M1 monoclonal antibody (b) was used to detect HA and M1, respectively.
    Rabbit Anti Ha Polyclonal Antibody, supplied by Sino Biological, 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/rabbit anti ha polyclonal antibody/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti ha polyclonal antibody - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

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    Western blot analysis of culture supernatant immunoprecipitated with rabbit anti-HA polyclonal antibody. Mouse anti-HA monoclonal antibody (a) and mouse anti-M1 monoclonal antibody (b) was used to detect HA and M1, respectively.

    Journal: Biochemical Engineering Journal

    Article Title: Production of influenza virus-like particles using recombinant insect cells

    doi: 10.1016/j.bej.2020.107757

    Figure Lengend Snippet: Western blot analysis of culture supernatant immunoprecipitated with rabbit anti-HA polyclonal antibody. Mouse anti-HA monoclonal antibody (a) and mouse anti-M1 monoclonal antibody (b) was used to detect HA and M1, respectively.

    Article Snippet: 2.7 ImmunoprecipitationCulture supernatant of recombinant cells was mixed with either rabbit anti-HA polyclonal antibody (Sino Biological) or rabbit anti-M1 polyclonal antibody (Sino Biological) in microtubes that were then incubated for 1 h. After protein G PLUS-agarose beads (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were added, the microtubes were rotated for 1 h. Bead-antigen complexes were then pelleted by centrifugation, and the pellet was washed 3 times with PBS.

    Techniques: Western Blot, Immunoprecipitation

    Biological and chemical modifications of H7N9 VLPs. (A) SDS-PAGE and western blot analysis of biologically modified H7N9 VLPs. Lane 1–3 represent 320 ng, 180 ng and 90 ng of αGal (-) VLP based on HA content, respectively. Lane 4–6 are 130 ng, 90, and 40 ng of αGal (+) VLP, respectively. (B) SDS-PAGE and western blot analysis of chemically modified H7N9 VLPs. S: protein standards; 1: αGal (-) H7N9 VLP; 2: intermediate of αGal (-) H7N9 VLP reacted with GO; 3: H7N9 conjugated with αGal linker a11; 4: H7N9 conjugated with αGal linker aN11; 5: H7N9 VLPs conjugated with control linker sp11. (C) Electron microscopy of H7N9 VLPs. (D) Electron microscopy of H7N9 VLPs stained with gold particles. Negative: VLPs were only stained with immunogold donkey anti-rabbit secondary antibody; Positive: VLPs were stained with anti-HA7 rabbit polyclonal antibody, followed by staining with immunogold donkey anti-rabbit secondary antibody. Sp11 is the aminooxy spacer control linker without αGal antigen; a11 is one type of αGal (Galα1,3-Galβ1,4-Glc) aminooxy linker; aN11 is the other type of αGal (Galα1,3-Galβ1,4-GlcNAc) aminooxy linker.

    Journal: PLoS ONE

    Article Title: Addition of αGal HyperAcute™ technology to recombinant avian influenza vaccines induces strong low-dose antibody responses

    doi: 10.1371/journal.pone.0182683

    Figure Lengend Snippet: Biological and chemical modifications of H7N9 VLPs. (A) SDS-PAGE and western blot analysis of biologically modified H7N9 VLPs. Lane 1–3 represent 320 ng, 180 ng and 90 ng of αGal (-) VLP based on HA content, respectively. Lane 4–6 are 130 ng, 90, and 40 ng of αGal (+) VLP, respectively. (B) SDS-PAGE and western blot analysis of chemically modified H7N9 VLPs. S: protein standards; 1: αGal (-) H7N9 VLP; 2: intermediate of αGal (-) H7N9 VLP reacted with GO; 3: H7N9 conjugated with αGal linker a11; 4: H7N9 conjugated with αGal linker aN11; 5: H7N9 VLPs conjugated with control linker sp11. (C) Electron microscopy of H7N9 VLPs. (D) Electron microscopy of H7N9 VLPs stained with gold particles. Negative: VLPs were only stained with immunogold donkey anti-rabbit secondary antibody; Positive: VLPs were stained with anti-HA7 rabbit polyclonal antibody, followed by staining with immunogold donkey anti-rabbit secondary antibody. Sp11 is the aminooxy spacer control linker without αGal antigen; a11 is one type of αGal (Galα1,3-Galβ1,4-Glc) aminooxy linker; aN11 is the other type of αGal (Galα1,3-Galβ1,4-GlcNAc) aminooxy linker.

    Article Snippet: Following a 30-minute incubation in a glass petri dish incubation chamber, the grid was stained in a 40 μL drop of rabbit polyclonal anti-HA7 antibody (Sino Biological Inc., 40103-RP02, 1: 500 dilution in blocking buffer) or blocking buffer alone to serve as a negative control.

    Techniques: SDS Page, Western Blot, Modification, Electron Microscopy, Staining

    M2 interacts with TRAPPC6A and TRAPPC6AΔ in mammalian cells. (A to C) Plasmids expressing TRAPPC6A-Myc and Flag-SC09M2 (A), TRAPPC6A-Myc and Flag-AH05M2 (B), or TRAPPC6A-Myc and Flag-WSNM2 (C) were transfected individually or in combination, as indicated, in HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Flag MAb or a mouse anti-Myc MAb and were subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (D) Western blotting of proteins bound to GST alone or to GST-TRAPPC6A. HEK293T cells transfected with pCAGGS-SC09M2 or with the pCAGGS vector were lysed with IP buffer, and the lysate was incubated with purified GST or GST-TRAPPC6A and then subjected to a pulldown assay. Equal volumes of proteins bound to the beads and the original cell lysates (5% of the input) were examined by Western blotting using a mouse anti-M2 MAb or a mouse anti-actin MAb, respectively. The GST-tagged proteins in the eluates were detected by Coomassie blue (CB) staining. (E) Confocal analysis of the distribution of M2 and TRAPPC6A proteins in A549 cells. pCAGGS-TRAPPC6A-myc and pCAGGS-Flag-SC09M2 were transfected individually or in combination into A549 cells and assessed by immunofluorescence staining. IAV M2 was detected with a mouse anti-Flag MAb and visualized with Alexa Fluor 488 (green). TRAPPC6A was detected with a rabbit anti-Myc polyclonal antibody and visualized with Alexa Fluor 546 (red). Yellow indicates colocalization of Alexa Fluor 546 and 488 in the merged image. (F) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1 or pEGFP-C1-BM2 into HEK293T cells for 48 h before the cells were lysed. Following immunoprecipitation of the cell lysates with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of BM2 and TRAPPC6A, respectively. (G) Co-IP of M2 and TRAPPC6AΔ. pCAGGS-Flag-SC09M2 was cotransfected with pCAGGS-TRAPPC6A-myc or pCAGGS-TRAPPC6AΔ-myc into HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Myc MAb and subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A or TRAPPC6AΔ, respectively.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: M2 interacts with TRAPPC6A and TRAPPC6AΔ in mammalian cells. (A to C) Plasmids expressing TRAPPC6A-Myc and Flag-SC09M2 (A), TRAPPC6A-Myc and Flag-AH05M2 (B), or TRAPPC6A-Myc and Flag-WSNM2 (C) were transfected individually or in combination, as indicated, in HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Flag MAb or a mouse anti-Myc MAb and were subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (D) Western blotting of proteins bound to GST alone or to GST-TRAPPC6A. HEK293T cells transfected with pCAGGS-SC09M2 or with the pCAGGS vector were lysed with IP buffer, and the lysate was incubated with purified GST or GST-TRAPPC6A and then subjected to a pulldown assay. Equal volumes of proteins bound to the beads and the original cell lysates (5% of the input) were examined by Western blotting using a mouse anti-M2 MAb or a mouse anti-actin MAb, respectively. The GST-tagged proteins in the eluates were detected by Coomassie blue (CB) staining. (E) Confocal analysis of the distribution of M2 and TRAPPC6A proteins in A549 cells. pCAGGS-TRAPPC6A-myc and pCAGGS-Flag-SC09M2 were transfected individually or in combination into A549 cells and assessed by immunofluorescence staining. IAV M2 was detected with a mouse anti-Flag MAb and visualized with Alexa Fluor 488 (green). TRAPPC6A was detected with a rabbit anti-Myc polyclonal antibody and visualized with Alexa Fluor 546 (red). Yellow indicates colocalization of Alexa Fluor 546 and 488 in the merged image. (F) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1 or pEGFP-C1-BM2 into HEK293T cells for 48 h before the cells were lysed. Following immunoprecipitation of the cell lysates with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of BM2 and TRAPPC6A, respectively. (G) Co-IP of M2 and TRAPPC6AΔ. pCAGGS-Flag-SC09M2 was cotransfected with pCAGGS-TRAPPC6A-myc or pCAGGS-TRAPPC6AΔ-myc into HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Myc MAb and subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A or TRAPPC6AΔ, respectively.

    Article Snippet: The collected cells were fixed with 4% PFA for 30 min and then stained for the surface expression of M2, HA, or FGF2 with the corresponding primary and secondary antibodies [mouse anti-M2 MAb 14C2 and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2, rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for HA, and rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for FGF2], without permeabilization treatment.

    Techniques: Expressing, Transfection, Immunoprecipitation, Western Blot, Plasmid Preparation, Incubation, Purification, Staining, Immunofluorescence, Co-Immunoprecipitation Assay

    A leucine residue at position 96 of M2 is required for the TRAPPC6A interaction. (A) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1, pEGFP-C1-SC09 M2, pEGFP-C1-SC09 M2EDTM, or pEGFP-C1-SC09 M2CT into HEK293T cells for 48 h before preparation for cell lysates. Following immunoprecipitation with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody and a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (B and C) Plasmids expressing TRAPPC6A-myc and Flag-SC09M2 or Flag-SC09M2 with different amino acid deletions in the C terminus were cotransfected into HEK293T cells for 48 h before the preparation of cell lysates. Following immunoprecipitation with a mouse anti-Flag MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: A leucine residue at position 96 of M2 is required for the TRAPPC6A interaction. (A) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1, pEGFP-C1-SC09 M2, pEGFP-C1-SC09 M2EDTM, or pEGFP-C1-SC09 M2CT into HEK293T cells for 48 h before preparation for cell lysates. Following immunoprecipitation with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody and a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (B and C) Plasmids expressing TRAPPC6A-myc and Flag-SC09M2 or Flag-SC09M2 with different amino acid deletions in the C terminus were cotransfected into HEK293T cells for 48 h before the preparation of cell lysates. Following immunoprecipitation with a mouse anti-Flag MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively.

    Article Snippet: The collected cells were fixed with 4% PFA for 30 min and then stained for the surface expression of M2, HA, or FGF2 with the corresponding primary and secondary antibodies [mouse anti-M2 MAb 14C2 and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2, rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for HA, and rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for FGF2], without permeabilization treatment.

    Techniques: Immunoprecipitation, Western Blot, Expressing

    Dynamics of the interaction of M2 and TRAPPC6AΔ in wt and mutant WSN virus-infected cells. A549 cells were infected with wt influenza virus WSN (A), or one of the M2 deletion mutants WSN M2Del1 (B) and WSN M2Del2 (C), at an MOI of 5. At 4, 6, 8, 10, and 14 h p.i., the infected cells were fixed and stained with mouse anti-M2 MAb 14C2 and rabbit anti-TRAPPC6A polyclonal antibody, followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG(H+L) (green) and Alexa Fluor 546 donkey anti-rabbit IgG(H+L) (red). Nuclei were stained with DAPI.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: Dynamics of the interaction of M2 and TRAPPC6AΔ in wt and mutant WSN virus-infected cells. A549 cells were infected with wt influenza virus WSN (A), or one of the M2 deletion mutants WSN M2Del1 (B) and WSN M2Del2 (C), at an MOI of 5. At 4, 6, 8, 10, and 14 h p.i., the infected cells were fixed and stained with mouse anti-M2 MAb 14C2 and rabbit anti-TRAPPC6A polyclonal antibody, followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG(H+L) (green) and Alexa Fluor 546 donkey anti-rabbit IgG(H+L) (red). Nuclei were stained with DAPI.

    Article Snippet: The collected cells were fixed with 4% PFA for 30 min and then stained for the surface expression of M2, HA, or FGF2 with the corresponding primary and secondary antibodies [mouse anti-M2 MAb 14C2 and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2, rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for HA, and rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for FGF2], without permeabilization treatment.

    Techniques: Mutagenesis, Infection, Staining, Incubation

    Mutation at position 96 of M2 affects its interaction with TRAPPC6A. (A) Sequence analysis of IAV M2 at position 96. All of the IAV M2 sequences deposited in GenBank by 6 July 2014 were downloaded. The identity of the amino acids at position 96 was statistically analyzed. (B) Plasmids expressing TRAPPC6A-Myc and Flag-WSNM2 or Flag-WSNM2 with different mutations at position 96 were cotransfected into HEK293T cells for 48 h before the preparation of cell lysates. Following immunoprecipitation with a mouse anti-Flag MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: Mutation at position 96 of M2 affects its interaction with TRAPPC6A. (A) Sequence analysis of IAV M2 at position 96. All of the IAV M2 sequences deposited in GenBank by 6 July 2014 were downloaded. The identity of the amino acids at position 96 was statistically analyzed. (B) Plasmids expressing TRAPPC6A-Myc and Flag-WSNM2 or Flag-WSNM2 with different mutations at position 96 were cotransfected into HEK293T cells for 48 h before the preparation of cell lysates. Following immunoprecipitation with a mouse anti-Flag MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively.

    Article Snippet: The collected cells were fixed with 4% PFA for 30 min and then stained for the surface expression of M2, HA, or FGF2 with the corresponding primary and secondary antibodies [mouse anti-M2 MAb 14C2 and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2, rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for HA, and rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for FGF2], without permeabilization treatment.

    Techniques: Mutagenesis, Sequencing, Expressing, Immunoprecipitation, Western Blot

    Effect of modulation of TRAPPC6AΔ expression on the cell surface expression of viral and cellular proteins. (A) A549 cells were transfected with siRNA targeting TRAPPC6AΔ or with nontargeting siRNA for 48 h and were then infected with the WSN virus at an MOI of 3. Cell lysates were processed at 8 and 10 h p.i. and subjected to Western blotting using a mouse anti-M2 MAb to detect the expression level of M2. (B) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. Cells were fixed at 8 and 10 h p.i., left nonpermeabilized, and stained with the mouse anti-M2 MAb and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2 surface expression analysis by flow cytometry. The graph shows the fluorescence intensity of M2 surface expression. (C) The TRAPPC6AΔ-overexpressing A549 cell line or the A549 control cell line transduced with an empty retrovirus was infected with the WSN virus at an MOI of 3. Cell lysates were processed at 8 and 10 h p.i. and subjected to Western blotting using a mouse anti-M2 MAb to detect the expression level of M2. (D) The TRAPPC6AΔ-overexpressing A549 cell line or the A549 control cell line transduced with an empty retrovirus was infected with the WSN virus at an MOI of 3. The cell surface expression of M2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B. (E) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. The cell surface expression of HA was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B by using the rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L). (F) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. The cell surface expression of FGF2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B by using the rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L). (G) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. At 2 h p.i., the culture medium was replaced with medium supplemented with 25 μM amantadine. The cell surface expression of M2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: Effect of modulation of TRAPPC6AΔ expression on the cell surface expression of viral and cellular proteins. (A) A549 cells were transfected with siRNA targeting TRAPPC6AΔ or with nontargeting siRNA for 48 h and were then infected with the WSN virus at an MOI of 3. Cell lysates were processed at 8 and 10 h p.i. and subjected to Western blotting using a mouse anti-M2 MAb to detect the expression level of M2. (B) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. Cells were fixed at 8 and 10 h p.i., left nonpermeabilized, and stained with the mouse anti-M2 MAb and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2 surface expression analysis by flow cytometry. The graph shows the fluorescence intensity of M2 surface expression. (C) The TRAPPC6AΔ-overexpressing A549 cell line or the A549 control cell line transduced with an empty retrovirus was infected with the WSN virus at an MOI of 3. Cell lysates were processed at 8 and 10 h p.i. and subjected to Western blotting using a mouse anti-M2 MAb to detect the expression level of M2. (D) The TRAPPC6AΔ-overexpressing A549 cell line or the A549 control cell line transduced with an empty retrovirus was infected with the WSN virus at an MOI of 3. The cell surface expression of M2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B. (E) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. The cell surface expression of HA was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B by using the rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L). (F) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. The cell surface expression of FGF2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B by using the rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L). (G) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. At 2 h p.i., the culture medium was replaced with medium supplemented with 25 μM amantadine. The cell surface expression of M2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B.

    Article Snippet: The collected cells were fixed with 4% PFA for 30 min and then stained for the surface expression of M2, HA, or FGF2 with the corresponding primary and secondary antibodies [mouse anti-M2 MAb 14C2 and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2, rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for HA, and rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for FGF2], without permeabilization treatment.

    Techniques: Expressing, Transfection, Infection, Western Blot, Staining, Flow Cytometry, Cytometry, Fluorescence, Transduction

    TRAPPC6AΔ positively modulates influenza virus infection. (A) Endogenous expression of TRAPPC6AΔ in A549 cells. Whole lysates of A549 cells grown in 12-well plates were subjected to Western blotting with a rabbit anti-TRAPPC6A polyclonal antibody. HEK293T cell lysates transiently transfected with pCAGGS-TRAPPC6A or pCAGGS-TRAPPC6AΔ were used as a control. (B) siRNA knockdown of TRAPPC6AΔ in A549 cells. A549 cells were transfected with siRNA targeting TRAPPC6AΔ or nontargeting siRNA for 48 h. Whole-cell lysates were then collected and analyzed by Western blotting with a rabbit anti-TRAPPC6A polyclonal antibody. (C) Cell viability of siRNA-treated A549 cells measured by using the CellTiter-Glo assay. A549 cells were transfected with siRNA as described above for panel B. The data are presented as means ± standard deviations for triplicate transfections. (D) Virus replication in siRNA-treated A549 cells. Cells transfected with siRNA as described above for panel B were infected with WSN virus. At 24 and 48 h p.i., supernatants were collected and titrated for infectious virus by plaque assays in MDCK cells. Three independent experiments were performed, and data are shown as means ± standard deviations for triplicates from a representative experiment. **, P

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: TRAPPC6AΔ positively modulates influenza virus infection. (A) Endogenous expression of TRAPPC6AΔ in A549 cells. Whole lysates of A549 cells grown in 12-well plates were subjected to Western blotting with a rabbit anti-TRAPPC6A polyclonal antibody. HEK293T cell lysates transiently transfected with pCAGGS-TRAPPC6A or pCAGGS-TRAPPC6AΔ were used as a control. (B) siRNA knockdown of TRAPPC6AΔ in A549 cells. A549 cells were transfected with siRNA targeting TRAPPC6AΔ or nontargeting siRNA for 48 h. Whole-cell lysates were then collected and analyzed by Western blotting with a rabbit anti-TRAPPC6A polyclonal antibody. (C) Cell viability of siRNA-treated A549 cells measured by using the CellTiter-Glo assay. A549 cells were transfected with siRNA as described above for panel B. The data are presented as means ± standard deviations for triplicate transfections. (D) Virus replication in siRNA-treated A549 cells. Cells transfected with siRNA as described above for panel B were infected with WSN virus. At 24 and 48 h p.i., supernatants were collected and titrated for infectious virus by plaque assays in MDCK cells. Three independent experiments were performed, and data are shown as means ± standard deviations for triplicates from a representative experiment. **, P

    Article Snippet: The collected cells were fixed with 4% PFA for 30 min and then stained for the surface expression of M2, HA, or FGF2 with the corresponding primary and secondary antibodies [mouse anti-M2 MAb 14C2 and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2, rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for HA, and rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for FGF2], without permeabilization treatment.

    Techniques: Infection, Expressing, Western Blot, Transfection, Glo Assay

    Confocal microscopy of WSN virus-infected cells stained for the Golgi apparatus or lysosomes. A549 cells were infected with the wt WSN virus at an MOI of 5. At the indicated time points, infected cells were fixed and stained with mouse anti-Giantin MAb and rabbit anti-M2 polyclonal antibody (A), mouse anti-LAMP1 MAb and rabbit anti-M2 polyclonal antibody (B), or mouse anti-LAMP1 MAb and rabbit anti-TRAPPC6A polyclonal antibody (C), followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG(H+L) (green) and Alexa Fluor 546 donkey anti-rabbit IgG(H+L) (red). Nuclei were stained with DAPI.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: Confocal microscopy of WSN virus-infected cells stained for the Golgi apparatus or lysosomes. A549 cells were infected with the wt WSN virus at an MOI of 5. At the indicated time points, infected cells were fixed and stained with mouse anti-Giantin MAb and rabbit anti-M2 polyclonal antibody (A), mouse anti-LAMP1 MAb and rabbit anti-M2 polyclonal antibody (B), or mouse anti-LAMP1 MAb and rabbit anti-TRAPPC6A polyclonal antibody (C), followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG(H+L) (green) and Alexa Fluor 546 donkey anti-rabbit IgG(H+L) (red). Nuclei were stained with DAPI.

    Article Snippet: The collected cells were fixed with 4% PFA for 30 min and then stained for the surface expression of M2, HA, or FGF2 with the corresponding primary and secondary antibodies [mouse anti-M2 MAb 14C2 and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2, rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for HA, and rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) for FGF2], without permeabilization treatment.

    Techniques: Confocal Microscopy, Infection, Staining, Incubation

    Expression and characterization of H7N9 VLP. a The viral titers of three recombinant baculoviruses were determined by plaque assay. The titer of recombinant baculovirus-HA (rBac-HA) was approxmiately 6 × 10 6 PFU/ml ( left ), the rBac-NA 3 × 10 7 PFU/ml (center), and the rBac-M1 1.1 × 10 8 PFU/ml ( right ). The protein expression of HA, NA, and M1 was identified respectively by immunofluorescence assay (lower panel). b Sf9 cells were co-infected with rBac-HA, rBac-NA, and rBac-M1 under various MOI combinations, and the VLPs were purified via sucrose gradient centrifugation. Formulation derived from the combinations of rBac-HA (MOI 2), rBac-NA (MOI 2), and rBac-M1 (MOI 2) co-infection exhibited the highest HA activity. c H7N9 VLP, H7/HA protein, N9/NA protein, and M1 proteins were detected by Western blot

    Journal: BMC Biotechnology

    Article Title: Multi-antigen avian influenza a (H7N9) virus-like particles: particulate characterizations and immunogenicity evaluation in murine and avian models

    doi: 10.1186/s12896-016-0321-6

    Figure Lengend Snippet: Expression and characterization of H7N9 VLP. a The viral titers of three recombinant baculoviruses were determined by plaque assay. The titer of recombinant baculovirus-HA (rBac-HA) was approxmiately 6 × 10 6 PFU/ml ( left ), the rBac-NA 3 × 10 7 PFU/ml (center), and the rBac-M1 1.1 × 10 8 PFU/ml ( right ). The protein expression of HA, NA, and M1 was identified respectively by immunofluorescence assay (lower panel). b Sf9 cells were co-infected with rBac-HA, rBac-NA, and rBac-M1 under various MOI combinations, and the VLPs were purified via sucrose gradient centrifugation. Formulation derived from the combinations of rBac-HA (MOI 2), rBac-NA (MOI 2), and rBac-M1 (MOI 2) co-infection exhibited the highest HA activity. c H7N9 VLP, H7/HA protein, N9/NA protein, and M1 proteins were detected by Western blot

    Article Snippet: Three days later, the monolayer of cells was washed and fixed with 80% acetone at −20 °C for 20 min. Rabbit polyclonal antibody against H7/HA peptide (Sino Biological, Beijing, China), rabbit polyclonal antibody against N9/NA peptide (ProSci, Poway, CA), and mouse polyclonal antibody against a recombinant M1 protein were applied at 1:10,000, 1:2,000, and 1:1,000 dilutions respectively and incubated for 1 h. After PBS wash, cells were further incubated with FITC-conjugated anti-rabbit or anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, PA).

    Techniques: Expressing, Recombinant, Plaque Assay, Immunofluorescence, Infection, Purification, Gradient Centrifugation, Derivative Assay, Activity Assay, Western Blot