mouse anti-flag Search Results


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  • 99
    Millipore m2 mouse antiflag antibody
    M2 Mouse Antiflag Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam mouse anti flag
    ZTL interacts directly with CHE. A, Yeast two-hybrid assays between ZTL and CHE or TOC1. FL, Full length; decoy, translationally fused LOV and Kelch repeat domains; LOV, LOV domain only; Kelch, Kelch repeat domain only. B, Coimmunoprecipitation experiments between CHE and ZTL performed in HEK293T cells. <t>GFP-CHE</t> or GFP alone was coexpressed with full-length or decoy ZTL translationally fused to a <t>FLAG</t> affinity tag. Immunoprecipitation (IP) was performed with the FLAG tag. IB, Immunoblot.
    Mouse Anti Flag, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Beyotime mouse anti flag
    ZTL interacts directly with CHE. A, Yeast two-hybrid assays between ZTL and CHE or TOC1. FL, Full length; decoy, translationally fused LOV and Kelch repeat domains; LOV, LOV domain only; Kelch, Kelch repeat domain only. B, Coimmunoprecipitation experiments between CHE and ZTL performed in HEK293T cells. <t>GFP-CHE</t> or GFP alone was coexpressed with full-length or decoy ZTL translationally fused to a <t>FLAG</t> affinity tag. Immunoprecipitation (IP) was performed with the FLAG tag. IB, Immunoblot.
    Mouse Anti Flag, supplied by Beyotime, used in various techniques. Bioz Stars score: 91/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ABM Inc mouse anti flag
    ZTL interacts directly with CHE. A, Yeast two-hybrid assays between ZTL and CHE or TOC1. FL, Full length; decoy, translationally fused LOV and Kelch repeat domains; LOV, LOV domain only; Kelch, Kelch repeat domain only. B, Coimmunoprecipitation experiments between CHE and ZTL performed in HEK293T cells. <t>GFP-CHE</t> or GFP alone was coexpressed with full-length or decoy ZTL translationally fused to a <t>FLAG</t> affinity tag. Immunoprecipitation (IP) was performed with the FLAG tag. IB, Immunoblot.
    Mouse Anti Flag, supplied by ABM Inc, used in various techniques. Bioz Stars score: 92/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abnova mouse anti flag
    ZTL interacts directly with CHE. A, Yeast two-hybrid assays between ZTL and CHE or TOC1. FL, Full length; decoy, translationally fused LOV and Kelch repeat domains; LOV, LOV domain only; Kelch, Kelch repeat domain only. B, Coimmunoprecipitation experiments between CHE and ZTL performed in HEK293T cells. <t>GFP-CHE</t> or GFP alone was coexpressed with full-length or decoy ZTL translationally fused to a <t>FLAG</t> affinity tag. Immunoprecipitation (IP) was performed with the FLAG tag. IB, Immunoblot.
    Mouse Anti Flag, supplied by Abnova, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Prospec mouse anti flag
    ZTL interacts directly with CHE. A, Yeast two-hybrid assays between ZTL and CHE or TOC1. FL, Full length; decoy, translationally fused LOV and Kelch repeat domains; LOV, LOV domain only; Kelch, Kelch repeat domain only. B, Coimmunoprecipitation experiments between CHE and ZTL performed in HEK293T cells. <t>GFP-CHE</t> or GFP alone was coexpressed with full-length or decoy ZTL translationally fused to a <t>FLAG</t> affinity tag. Immunoprecipitation (IP) was performed with the FLAG tag. IB, Immunoblot.
    Mouse Anti Flag, supplied by Prospec, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc mouse anti flag
    ZTL interacts directly with CHE. A, Yeast two-hybrid assays between ZTL and CHE or TOC1. FL, Full length; decoy, translationally fused LOV and Kelch repeat domains; LOV, LOV domain only; Kelch, Kelch repeat domain only. B, Coimmunoprecipitation experiments between CHE and ZTL performed in HEK293T cells. <t>GFP-CHE</t> or GFP alone was coexpressed with full-length or decoy ZTL translationally fused to a <t>FLAG</t> affinity tag. Immunoprecipitation (IP) was performed with the FLAG tag. IB, Immunoblot.
    Mouse Anti Flag, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 100 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Merck & Co mouse anti flag
    NEGR1 influences FGFR2 intracellular trafficking. ( A ) Representative immunoblots on protein extracts from lysates of HEK293 cells expressing control <t>FLAG-NSF</t> or FLAG-NEGR1, chemically cross-linked, solubilized and processed for FLAG-immunoprecipitation (pull down, FLAG-PD). NEGR1 interacts with FGFR2 but not with FGFR1, S6 ribosomial protein (S6rp) or ubiquitously expressed ATPase N -ethylmaleimide sensitive factor (NSF). INPUT, 10% of total lysate. Similar experiments were repeated four times. ( B ) Representative fluorescence images of FLAG and DAPI immunostainings in wild-type or FGFR2-expressing HEK293 cells (FGFR2) exposed to purified soluble FLAG-NEGR1 (sFLAG-Negr1) or first to sFLAG-Negr1 and subsequently to the pan-FGFRs activator FGFb. Scale bar = 20 μm. ( C ) Representative images of GFP fluorescence and DAPI staining in wild-type or N2A cells stably expressing FLAG-NEGR1 assayed for cluster-formation when transfected with GFP alone or FGFR2-GFP. Arrows indicate clustered cells; arrowheads indicate GFP-expressing cells. Scale bar = 20 μm. ( D ) Quantification of the number of cells organized in clusters in experiments as in C . Data are expressed as average number of cells organized in cluster. Asterisks indicate statistically significant difference (one-way ANOVA, post hoc Bonferroni test: *** P
    Mouse Anti Flag, supplied by Merck & Co, used in various techniques. Bioz Stars score: 93/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology mouse anti flag
    Activation of Raf-1 represses the association between Raf-1 and <t>STAT1.</t> (A) CA-STAT1 overexpressing or not Raf:ER cells were cultured either in the presence or absence 4HT for 48 h. Cell lysates were coimmunoprecipitated with the antibody to Raf-1 and then blotted for STAT1 or Raf-1 or <t>FLAG</t> tag on CA-STAT1 (a and b). (c) Raf:ER cell lysates in the absence of 4-HT were immunoprecipitated with Raf-1 or mouse IgG and analyzed by immunoblotting with STAT1 (lanes 1 and 2). Total lysates were immunoblotted for STAT1 as loading controls (lane 3 and 4). (B) Quantification of ERK activation. The ratios of phospho-ERK (green) to total ERK (red) were quantified (bottom) (mean ± SEM, n = 3). Activation of Raf-1 by 4HT induces ERK activation, yielding green and yellow bands. but CA-STAT1 overexpression has no effect on it.
    Mouse Anti Flag, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 115 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Agilent technologies mouse anti flag
    <t>Cdc25A</t> interacts directly with HDAC11 ( A ) and ( B ) Interaction between endogenous Cdc25A and HDAC11. Cdc25A (A) or HDAC11 (B) was immunoprecipitated from HEK 293T cell lysates. Immunoprecipitates were subjected to SDS-PAGE and probed with anti-HDAC11 and anti-Cdc25A antibodies. ( C ) Cdc25A binds directly to HDAC11. Purified His-HDAC11 and BSA (control) were immobilized on nitrocellulose membrane at the indicated concentrations and subsequently incubated in buffer containing <t>FLAG-Cdc25A</t> (800 ng). Cdc25A bound to His-HDAC11 was detected using anti-FLAG antibodies. ( D ) HDAC11 binds directly to Cdc25A. The reciprocal experiment used purified FLAG-Cdc25A and BSA immobilized on a nitrocellulose membrane which was incubated in buffer containing His-HDAC11 (800 ng). Bound His-HDAC11 was detected using anti-His antibodies.
    Mouse Anti Flag, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    TaKaRa mouse anti flag
    <t>Cdc25A</t> interacts directly with HDAC11 ( A ) and ( B ) Interaction between endogenous Cdc25A and HDAC11. Cdc25A (A) or HDAC11 (B) was immunoprecipitated from HEK 293T cell lysates. Immunoprecipitates were subjected to SDS-PAGE and probed with anti-HDAC11 and anti-Cdc25A antibodies. ( C ) Cdc25A binds directly to HDAC11. Purified His-HDAC11 and BSA (control) were immobilized on nitrocellulose membrane at the indicated concentrations and subsequently incubated in buffer containing <t>FLAG-Cdc25A</t> (800 ng). Cdc25A bound to His-HDAC11 was detected using anti-FLAG antibodies. ( D ) HDAC11 binds directly to Cdc25A. The reciprocal experiment used purified FLAG-Cdc25A and BSA immobilized on a nitrocellulose membrane which was incubated in buffer containing His-HDAC11 (800 ng). Bound His-HDAC11 was detected using anti-His antibodies.
    Mouse Anti Flag, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    OriGene mouse anti flag
    cGAS localizes in punctate regions around the chlamydial inclusion membrane HeLa cells infected with C. muridarum (A) or C. trachomatis , serovar D (B) were fixed and stained for endogenous cGAS (red) and Chlamydia (green). Cells were fixed with Prolong gold™ with DAPI (blue) and analyzed by confocal microscopy. Uninfected HeLa cells (C) and cells infected with C. muridarum (D) for 18 h were fixed and stained for endogenous cGAS (red) and STING (green). In an independent experiment HeLa cells were <t>transfected</t> with <t>FLAG-cGAS</t> (E) and infected with C. muridarum 24 h later. Infected cells were fixed at 18 h p.i and stained using mouse monoclonal Ab for FLAG. Chlamydial inclusions and cell nucleus are marked with an “I” and “N” respectively on the DAPI staining.
    Mouse Anti Flag, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Covance mouse anti flag
    cGAS localizes in punctate regions around the chlamydial inclusion membrane HeLa cells infected with C. muridarum (A) or C. trachomatis , serovar D (B) were fixed and stained for endogenous cGAS (red) and Chlamydia (green). Cells were fixed with Prolong gold™ with DAPI (blue) and analyzed by confocal microscopy. Uninfected HeLa cells (C) and cells infected with C. muridarum (D) for 18 h were fixed and stained for endogenous cGAS (red) and STING (green). In an independent experiment HeLa cells were <t>transfected</t> with <t>FLAG-cGAS</t> (E) and infected with C. muridarum 24 h later. Infected cells were fixed at 18 h p.i and stained using mouse monoclonal Ab for FLAG. Chlamydial inclusions and cell nucleus are marked with an “I” and “N” respectively on the DAPI staining.
    Mouse Anti Flag, supplied by Covance, used in various techniques. Bioz Stars score: 93/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    MBL International mouse anti flag
    cGAS localizes in punctate regions around the chlamydial inclusion membrane HeLa cells infected with C. muridarum (A) or C. trachomatis , serovar D (B) were fixed and stained for endogenous cGAS (red) and Chlamydia (green). Cells were fixed with Prolong gold™ with DAPI (blue) and analyzed by confocal microscopy. Uninfected HeLa cells (C) and cells infected with C. muridarum (D) for 18 h were fixed and stained for endogenous cGAS (red) and STING (green). In an independent experiment HeLa cells were <t>transfected</t> with <t>FLAG-cGAS</t> (E) and infected with C. muridarum 24 h later. Infected cells were fixed at 18 h p.i and stained using mouse monoclonal Ab for FLAG. Chlamydial inclusions and cell nucleus are marked with an “I” and “N” respectively on the DAPI staining.
    Mouse Anti Flag, supplied by MBL International, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    GenScript mouse anti flag
    Recombinant (RC) lubricin forms non-covalent complexes with collagen II and fibronectin. ( a ) Representation of RC lubricin constructs. Dark blue: <t>FLAG-tagged,</t> expressed in mammalian 293 F cells. Light blue: N-terminal fragments <t>GST-tagged</t> expressed in E. coli strain Rosetta 2. ( b ) Interaction between collagen II isolated from bovine cartilage and RC lubricin fragments, full length lubricin and BSA by solid phase binding assay. ( c ) Interaction between collagen II isolated from bovine cartilage and RC N-terminal lubricin fragments and BSA by solid phase binding assay. ( d ) Interaction between fibronectin isolated from human blood and RC lubricin fragments, full length lubricin and BSA by solid phase binding assay. For all assays n = 3 and error bars are standard deviation. N- designates the amino terminus and –C designates the carboxy terminus. The signal peptide (1–24) is shown in grey. All statistical analyses compare the binding of lubricin fragments to the binding of the BSA standard. * is defined as p ≤ 0.05, and *** is defined as p ≤ 0.001.
    Mouse Anti Flag, supplied by GenScript, used in various techniques. Bioz Stars score: 92/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Stratagene mouse anti flag
    Interaction between <t>ATF5</t> and HSP70. A , reciprocal immunoprecipitation ( IP ) analysis showing interaction between ATF5 and HSP70. The C6-pCIN4 and <t>C6-pCIN4-FLAG-HA-ATF5</t> cell lines were used for cell extract preparation. The expression of endogenous HSP70
    Mouse Anti Flag, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 85 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Syd Labs mouse anti flag
    Interaction between <t>ATF5</t> and HSP70. A , reciprocal immunoprecipitation ( IP ) analysis showing interaction between ATF5 and HSP70. The C6-pCIN4 and <t>C6-pCIN4-FLAG-HA-ATF5</t> cell lines were used for cell extract preparation. The expression of endogenous HSP70
    Mouse Anti Flag, supplied by Syd Labs, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Trans Genic inc mouse anti flag
    Interaction between <t>ATF5</t> and HSP70. A , reciprocal immunoprecipitation ( IP ) analysis showing interaction between ATF5 and HSP70. The C6-pCIN4 and <t>C6-pCIN4-FLAG-HA-ATF5</t> cell lines were used for cell extract preparation. The expression of endogenous HSP70
    Mouse Anti Flag, supplied by Trans Genic inc, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Roche mouse anti flag
    Interaction between <t>ATF5</t> and HSP70. A , reciprocal immunoprecipitation ( IP ) analysis showing interaction between ATF5 and HSP70. The C6-pCIN4 and <t>C6-pCIN4-FLAG-HA-ATF5</t> cell lines were used for cell extract preparation. The expression of endogenous HSP70
    Mouse Anti Flag, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher mouse flag
    <t>CacyBP</t> bound to C-terminal domains of V protein. (A) Identification of protein interaction partners of V protein by yeast two-hybrid screening of a CEF cDNA library. (B–D) Co-localization of CacyBP with V, VC, and VN proteins in DF-1 cells. DF-1 cells were plated on coverslips and transfected with <t>Flag-V,</t> Flag-VN and Flag-VC. Forty-eight hours after transfection, the cells were stained with mouse anti-FLAG and rabbit anti-CacyBP antibodies, which was followed by staining with donkey anti-rabbit Alexa Fluor® 488 (green) and goat anti-mouse Alexa Fluor ®; 594 (Red) as secondary antibodies. The nucleus was subsequently stained with Hoechst 33342, and the images were captured using an ANDOR Revolution WD confocal microscope. Pearson's coefficient were analysis by Imaris (Microscopy Image Analysis Software, Bitplane, Switzerland), and Pearson's coefficient > 0.5 were considered to be probable co-localization. (E–G) HEK-293T cells in 60 mm cell culture dishes were co-transfected with the pCAGEN-Flag-V (or Flag-VN or Flag-VC) and pCMV-HA-CacyBP expression plasmids. Transfected cells were harvested and lysed 48 h after transfection, and the experiment was conducted according to the manufacturer's instructions for the Co-Immunoprecipitation Kit. After washing, immunoprecipitated proteins were identified and analyzed by western blotting using anti-HA or anti-FLAG antibodies.
    Mouse Flag, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Campro Scientific mouse anti flag
    <t>CacyBP</t> bound to C-terminal domains of V protein. (A) Identification of protein interaction partners of V protein by yeast two-hybrid screening of a CEF cDNA library. (B–D) Co-localization of CacyBP with V, VC, and VN proteins in DF-1 cells. DF-1 cells were plated on coverslips and transfected with <t>Flag-V,</t> Flag-VN and Flag-VC. Forty-eight hours after transfection, the cells were stained with mouse anti-FLAG and rabbit anti-CacyBP antibodies, which was followed by staining with donkey anti-rabbit Alexa Fluor® 488 (green) and goat anti-mouse Alexa Fluor ®; 594 (Red) as secondary antibodies. The nucleus was subsequently stained with Hoechst 33342, and the images were captured using an ANDOR Revolution WD confocal microscope. Pearson's coefficient were analysis by Imaris (Microscopy Image Analysis Software, Bitplane, Switzerland), and Pearson's coefficient > 0.5 were considered to be probable co-localization. (E–G) HEK-293T cells in 60 mm cell culture dishes were co-transfected with the pCAGEN-Flag-V (or Flag-VN or Flag-VC) and pCMV-HA-CacyBP expression plasmids. Transfected cells were harvested and lysed 48 h after transfection, and the experiment was conducted according to the manufacturer's instructions for the Co-Immunoprecipitation Kit. After washing, immunoprecipitated proteins were identified and analyzed by western blotting using anti-HA or anti-FLAG antibodies.
    Mouse Anti Flag, supplied by Campro Scientific, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ZTL interacts directly with CHE. A, Yeast two-hybrid assays between ZTL and CHE or TOC1. FL, Full length; decoy, translationally fused LOV and Kelch repeat domains; LOV, LOV domain only; Kelch, Kelch repeat domain only. B, Coimmunoprecipitation experiments between CHE and ZTL performed in HEK293T cells. GFP-CHE or GFP alone was coexpressed with full-length or decoy ZTL translationally fused to a FLAG affinity tag. Immunoprecipitation (IP) was performed with the FLAG tag. IB, Immunoblot.

    Journal: Plant Physiology

    Article Title: Decoys Untangle Complicated Redundancy and Reveal Targets of Circadian Clock F-Box Proteins 1Decoys Untangle Complicated Redundancy and Reveal Targets of Circadian Clock F-Box Proteins 1 [OPEN]

    doi: 10.1104/pp.18.00331

    Figure Lengend Snippet: ZTL interacts directly with CHE. A, Yeast two-hybrid assays between ZTL and CHE or TOC1. FL, Full length; decoy, translationally fused LOV and Kelch repeat domains; LOV, LOV domain only; Kelch, Kelch repeat domain only. B, Coimmunoprecipitation experiments between CHE and ZTL performed in HEK293T cells. GFP-CHE or GFP alone was coexpressed with full-length or decoy ZTL translationally fused to a FLAG affinity tag. Immunoprecipitation (IP) was performed with the FLAG tag. IB, Immunoblot.

    Article Snippet: The cell lysate was diluted 3-fold with 50 m m sodium phosphate and incubated with 2 µg of USP2cc (Sigma-Aldrich, catalog no. U6653) at 37°C for 2 h. The mouse anti-FLAG and rabbit anti-GFP (Abcam, catalog no. Ab-290) antibodies were used to detect FLAG- or GFP-fused proteins, respectively.

    Techniques: Immunoprecipitation, FLAG-tag

    NEGR1 influences FGFR2 intracellular trafficking. ( A ) Representative immunoblots on protein extracts from lysates of HEK293 cells expressing control FLAG-NSF or FLAG-NEGR1, chemically cross-linked, solubilized and processed for FLAG-immunoprecipitation (pull down, FLAG-PD). NEGR1 interacts with FGFR2 but not with FGFR1, S6 ribosomial protein (S6rp) or ubiquitously expressed ATPase N -ethylmaleimide sensitive factor (NSF). INPUT, 10% of total lysate. Similar experiments were repeated four times. ( B ) Representative fluorescence images of FLAG and DAPI immunostainings in wild-type or FGFR2-expressing HEK293 cells (FGFR2) exposed to purified soluble FLAG-NEGR1 (sFLAG-Negr1) or first to sFLAG-Negr1 and subsequently to the pan-FGFRs activator FGFb. Scale bar = 20 μm. ( C ) Representative images of GFP fluorescence and DAPI staining in wild-type or N2A cells stably expressing FLAG-NEGR1 assayed for cluster-formation when transfected with GFP alone or FGFR2-GFP. Arrows indicate clustered cells; arrowheads indicate GFP-expressing cells. Scale bar = 20 μm. ( D ) Quantification of the number of cells organized in clusters in experiments as in C . Data are expressed as average number of cells organized in cluster. Asterisks indicate statistically significant difference (one-way ANOVA, post hoc Bonferroni test: *** P

    Journal: Brain

    Article Title: NEGR1 and FGFR2 cooperatively regulate cortical development and core behaviours related to autism disorders in mice

    doi: 10.1093/brain/awy190

    Figure Lengend Snippet: NEGR1 influences FGFR2 intracellular trafficking. ( A ) Representative immunoblots on protein extracts from lysates of HEK293 cells expressing control FLAG-NSF or FLAG-NEGR1, chemically cross-linked, solubilized and processed for FLAG-immunoprecipitation (pull down, FLAG-PD). NEGR1 interacts with FGFR2 but not with FGFR1, S6 ribosomial protein (S6rp) or ubiquitously expressed ATPase N -ethylmaleimide sensitive factor (NSF). INPUT, 10% of total lysate. Similar experiments were repeated four times. ( B ) Representative fluorescence images of FLAG and DAPI immunostainings in wild-type or FGFR2-expressing HEK293 cells (FGFR2) exposed to purified soluble FLAG-NEGR1 (sFLAG-Negr1) or first to sFLAG-Negr1 and subsequently to the pan-FGFRs activator FGFb. Scale bar = 20 μm. ( C ) Representative images of GFP fluorescence and DAPI staining in wild-type or N2A cells stably expressing FLAG-NEGR1 assayed for cluster-formation when transfected with GFP alone or FGFR2-GFP. Arrows indicate clustered cells; arrowheads indicate GFP-expressing cells. Scale bar = 20 μm. ( D ) Quantification of the number of cells organized in clusters in experiments as in C . Data are expressed as average number of cells organized in cluster. Asterisks indicate statistically significant difference (one-way ANOVA, post hoc Bonferroni test: *** P

    Article Snippet: The primary antibodies were applied overnight in a blocking buffer (20 mM Tris, pH 7.4, 150 mM NaCl, 0.1% Tween 20, and 5% non-fat dried milk); primary antibodies included rabbit anti-FGFR2 1:200 (Santa Cruz Biotechnology), rabbit anti-FGFR1 1:200 (Santa Cruz Biotechnology), goat anti-Negr1 1:1000 (R & D), mouse anti-Na+ K+ ATPase (Abcam), rabbit anti-GFP 1:5000 (Life Technologies), rabbit anti-S6rp 1:2000, rabbit anti-p42/44 (pERK), rabbit anti-42/44 (ERK; Cell Signaling), rabbit anti-pAKT (Cell Signaling), rabbit anti-NSF (Cell Signaling), mouse anti-AKT pan (Cell Signaling), mouse anti-FLAG (Merck) and mouse anti-β-tubulin (Merck).

    Techniques: Western Blot, Expressing, Immunoprecipitation, Fluorescence, Purification, Staining, Stable Transfection, Transfection

    Activation of Raf-1 represses the association between Raf-1 and STAT1. (A) CA-STAT1 overexpressing or not Raf:ER cells were cultured either in the presence or absence 4HT for 48 h. Cell lysates were coimmunoprecipitated with the antibody to Raf-1 and then blotted for STAT1 or Raf-1 or FLAG tag on CA-STAT1 (a and b). (c) Raf:ER cell lysates in the absence of 4-HT were immunoprecipitated with Raf-1 or mouse IgG and analyzed by immunoblotting with STAT1 (lanes 1 and 2). Total lysates were immunoblotted for STAT1 as loading controls (lane 3 and 4). (B) Quantification of ERK activation. The ratios of phospho-ERK (green) to total ERK (red) were quantified (bottom) (mean ± SEM, n = 3). Activation of Raf-1 by 4HT induces ERK activation, yielding green and yellow bands. but CA-STAT1 overexpression has no effect on it.

    Journal: Molecular Biology of the Cell

    Article Title: STAT1 Is Required for Redifferentiation during Madin-Darby Canine Kidney Tubulogenesis

    doi: 10.1091/mbc.E10-02-0112

    Figure Lengend Snippet: Activation of Raf-1 represses the association between Raf-1 and STAT1. (A) CA-STAT1 overexpressing or not Raf:ER cells were cultured either in the presence or absence 4HT for 48 h. Cell lysates were coimmunoprecipitated with the antibody to Raf-1 and then blotted for STAT1 or Raf-1 or FLAG tag on CA-STAT1 (a and b). (c) Raf:ER cell lysates in the absence of 4-HT were immunoprecipitated with Raf-1 or mouse IgG and analyzed by immunoblotting with STAT1 (lanes 1 and 2). Total lysates were immunoblotted for STAT1 as loading controls (lane 3 and 4). (B) Quantification of ERK activation. The ratios of phospho-ERK (green) to total ERK (red) were quantified (bottom) (mean ± SEM, n = 3). Activation of Raf-1 by 4HT induces ERK activation, yielding green and yellow bands. but CA-STAT1 overexpression has no effect on it.

    Article Snippet: Primary antibodies were mouse anti-STAT1, mouse anti-phospho-STAT1 (Y701), mouse anti-Raf-1 (BD Biosciences, San Jose, CA); mouse anti-STAT1, rabbit anti-phospho-STAT1 (Y701) (Cell Signaling Technology, Danvers, MA); rabbit anti-ERK1/2, mouse anti-phosphoERK1/2, mouse anti-FLAG (Santa Cruz Biotechnology, Santa Cruz, CA); and mouse anti-glyceraldehyde-3-phosphate dehydrogenase ([GAPDH] Millipore Bioscience Research Reagents, Temecula, CA).

    Techniques: Activation Assay, Cell Culture, FLAG-tag, Immunoprecipitation, Over Expression

    Overexpression of tyrosine phosphorylation defective STAT1 partially rescues cord formation of activated Raf:ER cysts. (A) Raf:ER cysts (left) and three clones overexpressing Y701F-STAT1 were treated with 4HT for 72 h. Bar, 10 μm. (B) Total lysate from Raf:ER cells and three Y701F-STAT1 transfectants were immunoblotted for FLAG tag on Y701F-STAT1. (C) Quantification of cords in the presence of 4HT (mean ± SEM, n = 3). *p

    Journal: Molecular Biology of the Cell

    Article Title: STAT1 Is Required for Redifferentiation during Madin-Darby Canine Kidney Tubulogenesis

    doi: 10.1091/mbc.E10-02-0112

    Figure Lengend Snippet: Overexpression of tyrosine phosphorylation defective STAT1 partially rescues cord formation of activated Raf:ER cysts. (A) Raf:ER cysts (left) and three clones overexpressing Y701F-STAT1 were treated with 4HT for 72 h. Bar, 10 μm. (B) Total lysate from Raf:ER cells and three Y701F-STAT1 transfectants were immunoblotted for FLAG tag on Y701F-STAT1. (C) Quantification of cords in the presence of 4HT (mean ± SEM, n = 3). *p

    Article Snippet: Primary antibodies were mouse anti-STAT1, mouse anti-phospho-STAT1 (Y701), mouse anti-Raf-1 (BD Biosciences, San Jose, CA); mouse anti-STAT1, rabbit anti-phospho-STAT1 (Y701) (Cell Signaling Technology, Danvers, MA); rabbit anti-ERK1/2, mouse anti-phosphoERK1/2, mouse anti-FLAG (Santa Cruz Biotechnology, Santa Cruz, CA); and mouse anti-glyceraldehyde-3-phosphate dehydrogenase ([GAPDH] Millipore Bioscience Research Reagents, Temecula, CA).

    Techniques: Over Expression, Clone Assay, FLAG-tag

    Cdc25A interacts directly with HDAC11 ( A ) and ( B ) Interaction between endogenous Cdc25A and HDAC11. Cdc25A (A) or HDAC11 (B) was immunoprecipitated from HEK 293T cell lysates. Immunoprecipitates were subjected to SDS-PAGE and probed with anti-HDAC11 and anti-Cdc25A antibodies. ( C ) Cdc25A binds directly to HDAC11. Purified His-HDAC11 and BSA (control) were immobilized on nitrocellulose membrane at the indicated concentrations and subsequently incubated in buffer containing FLAG-Cdc25A (800 ng). Cdc25A bound to His-HDAC11 was detected using anti-FLAG antibodies. ( D ) HDAC11 binds directly to Cdc25A. The reciprocal experiment used purified FLAG-Cdc25A and BSA immobilized on a nitrocellulose membrane which was incubated in buffer containing His-HDAC11 (800 ng). Bound His-HDAC11 was detected using anti-His antibodies.

    Journal: Oncotarget

    Article Title: Acetylation and deacetylation of Cdc25A constitutes a novel mechanism for modulating Cdc25A functions with implications for cancer

    doi: 10.18632/oncotarget.7966

    Figure Lengend Snippet: Cdc25A interacts directly with HDAC11 ( A ) and ( B ) Interaction between endogenous Cdc25A and HDAC11. Cdc25A (A) or HDAC11 (B) was immunoprecipitated from HEK 293T cell lysates. Immunoprecipitates were subjected to SDS-PAGE and probed with anti-HDAC11 and anti-Cdc25A antibodies. ( C ) Cdc25A binds directly to HDAC11. Purified His-HDAC11 and BSA (control) were immobilized on nitrocellulose membrane at the indicated concentrations and subsequently incubated in buffer containing FLAG-Cdc25A (800 ng). Cdc25A bound to His-HDAC11 was detected using anti-FLAG antibodies. ( D ) HDAC11 binds directly to Cdc25A. The reciprocal experiment used purified FLAG-Cdc25A and BSA immobilized on a nitrocellulose membrane which was incubated in buffer containing His-HDAC11 (800 ng). Bound His-HDAC11 was detected using anti-His antibodies.

    Article Snippet: Membranes were washed 3 times, for 10 min each, with TBST, and were subjected to immunodetection with mouse anti-Cdc25A (Santa Cruz Biotechnology), mouse anti-GFP (Jackson ImmunoResearch), mouse anti-FLAG (Agilent Technologies), or mouse anti-His (Oncogene) antibodies for 1 h at 4°C (see figure legends for specifics of each experiment).

    Techniques: Immunoprecipitation, SDS Page, Purification, Incubation

    ARD1 acetylates Cdc25A in vitro and in cells ( A ) ARD1 mediates Cdc25A acetylation in vitro . Purified GFP-ARD1A was incubated at 4°C, or 37°C, with purified FLAG-Cdc25A. The reactions were analyzed using anti-acetyl lysine antibody, followed by chemiluminescent detection. ( B ) Cdc25A is endogenously acetylated. Cdc25A was immunoprecipitated from HEK 293T cell lysates and its acetylation status was analyzed by Western blot using an acetyl-specific antibody. ( C ) ARD1 overexpression increases Cdc25A acetylation in cells. Cdc25A was immunoprecipitated from HEK 293T cell lysates transfected with FLAG-ARD1A and the Cdc25A acetylation status was analyzed by Western blot using an antibody specific for acetylated lysines.

    Journal: Oncotarget

    Article Title: Acetylation and deacetylation of Cdc25A constitutes a novel mechanism for modulating Cdc25A functions with implications for cancer

    doi: 10.18632/oncotarget.7966

    Figure Lengend Snippet: ARD1 acetylates Cdc25A in vitro and in cells ( A ) ARD1 mediates Cdc25A acetylation in vitro . Purified GFP-ARD1A was incubated at 4°C, or 37°C, with purified FLAG-Cdc25A. The reactions were analyzed using anti-acetyl lysine antibody, followed by chemiluminescent detection. ( B ) Cdc25A is endogenously acetylated. Cdc25A was immunoprecipitated from HEK 293T cell lysates and its acetylation status was analyzed by Western blot using an acetyl-specific antibody. ( C ) ARD1 overexpression increases Cdc25A acetylation in cells. Cdc25A was immunoprecipitated from HEK 293T cell lysates transfected with FLAG-ARD1A and the Cdc25A acetylation status was analyzed by Western blot using an antibody specific for acetylated lysines.

    Article Snippet: Membranes were washed 3 times, for 10 min each, with TBST, and were subjected to immunodetection with mouse anti-Cdc25A (Santa Cruz Biotechnology), mouse anti-GFP (Jackson ImmunoResearch), mouse anti-FLAG (Agilent Technologies), or mouse anti-His (Oncogene) antibodies for 1 h at 4°C (see figure legends for specifics of each experiment).

    Techniques: In Vitro, Purification, Incubation, Immunoprecipitation, Western Blot, Over Expression, Transfection

    Cdc25A and ARD1 interact in vivo and in vitro ( A ) Cdc25A co-immunoprecipitates with ARD1 in vitro . Purified FLAG-Cdc25A was incubated with anti-GFP beads, either alone or in combination with GFP-ARD1A. Immunoprecipitated products were subjected to SDS-PAGE and probed with anti-GFP and anti-FLAG antibodies. ( B ) ARD1 and Cdc25A associate after co-transfection. FLAG-ARD1A was co-transfected into HEK 293T cells with either GFP-Cdc25A or the GFP vector alone. After 24 hours, cell lysates were subjected to immunoprecipitation using anti-GFP antibody. Precipitated proteins were examined by Western blot. ( C ) Co-immunoprecipitation of Cdc25A and ARD1 in cultured cells. HEK 293T cell lysates were subjected to immunoprecipitation using antibody to Cdc25A or mouse IgG. Immunoprecipitates were analyzed with anti-ARD1 and anti-Cdc25A antibodies. ( D ) and ( E ) Cdc25A-ARD1 interact directly. Purified ARD1A (D), Cdc25A (E), and a BSA control were immobilized on nitrocellulose membranes at the concentrations indicated above each panel. Membranes were incubated in buffer containing (D) Cdc25A (2400 ng) or (E) ARD1A (720 ng). Immunodetection of bound protein was performed with anti-Cdc25A (D) or anti-GFP (E) antibodies.

    Journal: Oncotarget

    Article Title: Acetylation and deacetylation of Cdc25A constitutes a novel mechanism for modulating Cdc25A functions with implications for cancer

    doi: 10.18632/oncotarget.7966

    Figure Lengend Snippet: Cdc25A and ARD1 interact in vivo and in vitro ( A ) Cdc25A co-immunoprecipitates with ARD1 in vitro . Purified FLAG-Cdc25A was incubated with anti-GFP beads, either alone or in combination with GFP-ARD1A. Immunoprecipitated products were subjected to SDS-PAGE and probed with anti-GFP and anti-FLAG antibodies. ( B ) ARD1 and Cdc25A associate after co-transfection. FLAG-ARD1A was co-transfected into HEK 293T cells with either GFP-Cdc25A or the GFP vector alone. After 24 hours, cell lysates were subjected to immunoprecipitation using anti-GFP antibody. Precipitated proteins were examined by Western blot. ( C ) Co-immunoprecipitation of Cdc25A and ARD1 in cultured cells. HEK 293T cell lysates were subjected to immunoprecipitation using antibody to Cdc25A or mouse IgG. Immunoprecipitates were analyzed with anti-ARD1 and anti-Cdc25A antibodies. ( D ) and ( E ) Cdc25A-ARD1 interact directly. Purified ARD1A (D), Cdc25A (E), and a BSA control were immobilized on nitrocellulose membranes at the concentrations indicated above each panel. Membranes were incubated in buffer containing (D) Cdc25A (2400 ng) or (E) ARD1A (720 ng). Immunodetection of bound protein was performed with anti-Cdc25A (D) or anti-GFP (E) antibodies.

    Article Snippet: Membranes were washed 3 times, for 10 min each, with TBST, and were subjected to immunodetection with mouse anti-Cdc25A (Santa Cruz Biotechnology), mouse anti-GFP (Jackson ImmunoResearch), mouse anti-FLAG (Agilent Technologies), or mouse anti-His (Oncogene) antibodies for 1 h at 4°C (see figure legends for specifics of each experiment).

    Techniques: In Vivo, In Vitro, Purification, Incubation, Immunoprecipitation, SDS Page, Cotransfection, Transfection, Plasmid Preparation, Western Blot, Cell Culture, Immunodetection

    cGAS localizes in punctate regions around the chlamydial inclusion membrane HeLa cells infected with C. muridarum (A) or C. trachomatis , serovar D (B) were fixed and stained for endogenous cGAS (red) and Chlamydia (green). Cells were fixed with Prolong gold™ with DAPI (blue) and analyzed by confocal microscopy. Uninfected HeLa cells (C) and cells infected with C. muridarum (D) for 18 h were fixed and stained for endogenous cGAS (red) and STING (green). In an independent experiment HeLa cells were transfected with FLAG-cGAS (E) and infected with C. muridarum 24 h later. Infected cells were fixed at 18 h p.i and stained using mouse monoclonal Ab for FLAG. Chlamydial inclusions and cell nucleus are marked with an “I” and “N” respectively on the DAPI staining.

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: The DNA sensor, cyclic GMP-AMP synthase (cGAS) is essential for induction of IFN beta during Chlamydia trachomatis infection

    doi: 10.4049/jimmunol.1302718

    Figure Lengend Snippet: cGAS localizes in punctate regions around the chlamydial inclusion membrane HeLa cells infected with C. muridarum (A) or C. trachomatis , serovar D (B) were fixed and stained for endogenous cGAS (red) and Chlamydia (green). Cells were fixed with Prolong gold™ with DAPI (blue) and analyzed by confocal microscopy. Uninfected HeLa cells (C) and cells infected with C. muridarum (D) for 18 h were fixed and stained for endogenous cGAS (red) and STING (green). In an independent experiment HeLa cells were transfected with FLAG-cGAS (E) and infected with C. muridarum 24 h later. Infected cells were fixed at 18 h p.i and stained using mouse monoclonal Ab for FLAG. Chlamydial inclusions and cell nucleus are marked with an “I” and “N” respectively on the DAPI staining.

    Article Snippet: Transfected cells were stained using mouse anti-FLAG (Origene).

    Techniques: Infection, Staining, Confocal Microscopy, Transfection

    Recombinant (RC) lubricin forms non-covalent complexes with collagen II and fibronectin. ( a ) Representation of RC lubricin constructs. Dark blue: FLAG-tagged, expressed in mammalian 293 F cells. Light blue: N-terminal fragments GST-tagged expressed in E. coli strain Rosetta 2. ( b ) Interaction between collagen II isolated from bovine cartilage and RC lubricin fragments, full length lubricin and BSA by solid phase binding assay. ( c ) Interaction between collagen II isolated from bovine cartilage and RC N-terminal lubricin fragments and BSA by solid phase binding assay. ( d ) Interaction between fibronectin isolated from human blood and RC lubricin fragments, full length lubricin and BSA by solid phase binding assay. For all assays n = 3 and error bars are standard deviation. N- designates the amino terminus and –C designates the carboxy terminus. The signal peptide (1–24) is shown in grey. All statistical analyses compare the binding of lubricin fragments to the binding of the BSA standard. * is defined as p ≤ 0.05, and *** is defined as p ≤ 0.001.

    Journal: Scientific Reports

    Article Title: Lubricin binds cartilage proteins, cartilage oligomeric matrix protein, fibronectin and collagen II at the cartilage surface

    doi: 10.1038/s41598-017-13558-y

    Figure Lengend Snippet: Recombinant (RC) lubricin forms non-covalent complexes with collagen II and fibronectin. ( a ) Representation of RC lubricin constructs. Dark blue: FLAG-tagged, expressed in mammalian 293 F cells. Light blue: N-terminal fragments GST-tagged expressed in E. coli strain Rosetta 2. ( b ) Interaction between collagen II isolated from bovine cartilage and RC lubricin fragments, full length lubricin and BSA by solid phase binding assay. ( c ) Interaction between collagen II isolated from bovine cartilage and RC N-terminal lubricin fragments and BSA by solid phase binding assay. ( d ) Interaction between fibronectin isolated from human blood and RC lubricin fragments, full length lubricin and BSA by solid phase binding assay. For all assays n = 3 and error bars are standard deviation. N- designates the amino terminus and –C designates the carboxy terminus. The signal peptide (1–24) is shown in grey. All statistical analyses compare the binding of lubricin fragments to the binding of the BSA standard. * is defined as p ≤ 0.05, and *** is defined as p ≤ 0.001.

    Article Snippet: After a TBS rinse, the plates were incubated with lubricin fragments at 2 µg/ml in wash buffer (TBS + 0.1% BSA + 0.1% Tween-20) or BSA as a negative control, for 2 h. Plates were then washed three times with wash buffer and incubated with mouse anti-FLAG (Genscript) or rabbit anti-GST (Abcam) antibodies at 1 µg/ml in wash buffer for 1 h. The plates were then incubated with anti-mouse or anti-rabbit HRP-conjugated antibodies (DAKO) at 0.2 µg/ml in wash buffer for 1 h. After washing, binding was detected using TMB substrate (ThermoFisher), signal stopped with sulphuric acid and absorbance read at 450 nm.

    Techniques: Recombinant, Construct, Isolation, Binding Assay, Standard Deviation

    Interaction between ATF5 and HSP70. A , reciprocal immunoprecipitation ( IP ) analysis showing interaction between ATF5 and HSP70. The C6-pCIN4 and C6-pCIN4-FLAG-HA-ATF5 cell lines were used for cell extract preparation. The expression of endogenous HSP70

    Journal: The Journal of Biological Chemistry

    Article Title: HSP70 Protein Promotes Survival of C6 and U87 Glioma Cells by Inhibition of ATF5 Degradation *

    doi: 10.1074/jbc.M110.211771

    Figure Lengend Snippet: Interaction between ATF5 and HSP70. A , reciprocal immunoprecipitation ( IP ) analysis showing interaction between ATF5 and HSP70. The C6-pCIN4 and C6-pCIN4-FLAG-HA-ATF5 cell lines were used for cell extract preparation. The expression of endogenous HSP70

    Article Snippet: The antibodies used for immunoblotting analysis were rabbit anti-ATF5 (1:250) and mouse anti-β-actin (1:10,000) from Abcam; mouse anti-FLAG (1:1000) from Stratagene; rat anti-HA (1:1,000) from Roche; rabbit anti-Egr-1 (1:200), rabbit anti-HSP70 (1:200), B-Raf (1:100), and anti-HSP90 (1:200) antibodies from Santa Cruz Biotechnology; and mouse anti-Bcl-2 (1:250) from BD Pharmingen.

    Techniques: Immunoprecipitation, Expressing

    CacyBP bound to C-terminal domains of V protein. (A) Identification of protein interaction partners of V protein by yeast two-hybrid screening of a CEF cDNA library. (B–D) Co-localization of CacyBP with V, VC, and VN proteins in DF-1 cells. DF-1 cells were plated on coverslips and transfected with Flag-V, Flag-VN and Flag-VC. Forty-eight hours after transfection, the cells were stained with mouse anti-FLAG and rabbit anti-CacyBP antibodies, which was followed by staining with donkey anti-rabbit Alexa Fluor® 488 (green) and goat anti-mouse Alexa Fluor ®; 594 (Red) as secondary antibodies. The nucleus was subsequently stained with Hoechst 33342, and the images were captured using an ANDOR Revolution WD confocal microscope. Pearson's coefficient were analysis by Imaris (Microscopy Image Analysis Software, Bitplane, Switzerland), and Pearson's coefficient > 0.5 were considered to be probable co-localization. (E–G) HEK-293T cells in 60 mm cell culture dishes were co-transfected with the pCAGEN-Flag-V (or Flag-VN or Flag-VC) and pCMV-HA-CacyBP expression plasmids. Transfected cells were harvested and lysed 48 h after transfection, and the experiment was conducted according to the manufacturer's instructions for the Co-Immunoprecipitation Kit. After washing, immunoprecipitated proteins were identified and analyzed by western blotting using anti-HA or anti-FLAG antibodies.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Newcastle Disease Virus V Protein Inhibits Cell Apoptosis and Promotes Viral Replication by Targeting CacyBP/SIP

    doi: 10.3389/fcimb.2018.00304

    Figure Lengend Snippet: CacyBP bound to C-terminal domains of V protein. (A) Identification of protein interaction partners of V protein by yeast two-hybrid screening of a CEF cDNA library. (B–D) Co-localization of CacyBP with V, VC, and VN proteins in DF-1 cells. DF-1 cells were plated on coverslips and transfected with Flag-V, Flag-VN and Flag-VC. Forty-eight hours after transfection, the cells were stained with mouse anti-FLAG and rabbit anti-CacyBP antibodies, which was followed by staining with donkey anti-rabbit Alexa Fluor® 488 (green) and goat anti-mouse Alexa Fluor ®; 594 (Red) as secondary antibodies. The nucleus was subsequently stained with Hoechst 33342, and the images were captured using an ANDOR Revolution WD confocal microscope. Pearson's coefficient were analysis by Imaris (Microscopy Image Analysis Software, Bitplane, Switzerland), and Pearson's coefficient > 0.5 were considered to be probable co-localization. (E–G) HEK-293T cells in 60 mm cell culture dishes were co-transfected with the pCAGEN-Flag-V (or Flag-VN or Flag-VC) and pCMV-HA-CacyBP expression plasmids. Transfected cells were harvested and lysed 48 h after transfection, and the experiment was conducted according to the manufacturer's instructions for the Co-Immunoprecipitation Kit. After washing, immunoprecipitated proteins were identified and analyzed by western blotting using anti-HA or anti-FLAG antibodies.

    Article Snippet: The specific primary antibodies of rabbit anti-CacyBP/SIP (1:100, BOSTER, China), mouse anti-FLAG (1:500, Invitrogen, USA) or mouse anti-HA (1:500, Invitrogen, USA) were used as appropriate.

    Techniques: Two Hybrid Screening, cDNA Library Assay, Transfection, Staining, Microscopy, Software, Cell Culture, Expressing, Immunoprecipitation, Western Blot

    V protein inhibited apoptosis by downregulating CacyBP in DF-1 cells. DF-1 cells were transfected with the control (pCAGEN-Flag), pCAGEN-Flag-VC, and pCAGEN-Flag-V. (A) The transfected cells were washed and harvested 48 h after transfection, and flow cytometry was used to analyze cell apoptosis. (B) Twenty-four and forty-eight hours after transfection, the mRNA levels of CacyBP were measured by Q-PCR. (C) Protein expression of CacyBP was measured in DF-1 cells. The cells were transfected with pCAGEN-Flag (control) and pCAGEN-Flag-V (1, 2, and 4 μg), and after 48 h, the cells were harvested and analyzed by western blotting. Data shown in (A,B) are mean ± SD of three independent experiments. * P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Newcastle Disease Virus V Protein Inhibits Cell Apoptosis and Promotes Viral Replication by Targeting CacyBP/SIP

    doi: 10.3389/fcimb.2018.00304

    Figure Lengend Snippet: V protein inhibited apoptosis by downregulating CacyBP in DF-1 cells. DF-1 cells were transfected with the control (pCAGEN-Flag), pCAGEN-Flag-VC, and pCAGEN-Flag-V. (A) The transfected cells were washed and harvested 48 h after transfection, and flow cytometry was used to analyze cell apoptosis. (B) Twenty-four and forty-eight hours after transfection, the mRNA levels of CacyBP were measured by Q-PCR. (C) Protein expression of CacyBP was measured in DF-1 cells. The cells were transfected with pCAGEN-Flag (control) and pCAGEN-Flag-V (1, 2, and 4 μg), and after 48 h, the cells were harvested and analyzed by western blotting. Data shown in (A,B) are mean ± SD of three independent experiments. * P

    Article Snippet: The specific primary antibodies of rabbit anti-CacyBP/SIP (1:100, BOSTER, China), mouse anti-FLAG (1:500, Invitrogen, USA) or mouse anti-HA (1:500, Invitrogen, USA) were used as appropriate.

    Techniques: Transfection, Flow Cytometry, Cytometry, Polymerase Chain Reaction, Expressing, Western Blot