bl21 de3 cells Thermo Fisher Search Results


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  • 99
    Thermo Fisher bl21 de3 competent cells
    Bl21 De3 Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 127 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher bl21 cells
    SDS-PAGE and western blot analysis of E. coli <t>BL21-AI</t> cells expressing B. melitensis Omp31, BP26, or 16. Panel A. SDS-PAGE analysis of E. coli BL21-AI cells harbouring pDESTOMPs of B. melitensis. (M) Marker lane, (1) uninduced, (2), (3) and (4) are Omp31, BP26, and Omp16, respectively. Panel B. Western blot analysis of induced samples using anti-HIS probe. Lanes (1), (2) and (3) are B. melitensis Omp31, BP26, and Omp16, respectively.
    Bl21 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 404 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bl21 de3 cells
    SDS-PAGE and western blot analysis of E. coli <t>BL21-AI</t> cells expressing B. melitensis Omp31, BP26, or 16. Panel A. SDS-PAGE analysis of E. coli BL21-AI cells harbouring pDESTOMPs of B. melitensis. (M) Marker lane, (1) uninduced, (2), (3) and (4) are Omp31, BP26, and Omp16, respectively. Panel B. Western blot analysis of induced samples using anti-HIS probe. Lanes (1), (2) and (3) are B. melitensis Omp31, BP26, and Omp16, respectively.
    Bl21 De3 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1125 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher competent e coli cells
    SDS-PAGE and western blot analysis of E. coli <t>BL21-AI</t> cells expressing B. melitensis Omp31, BP26, or 16. Panel A. SDS-PAGE analysis of E. coli BL21-AI cells harbouring pDESTOMPs of B. melitensis. (M) Marker lane, (1) uninduced, (2), (3) and (4) are Omp31, BP26, and Omp16, respectively. Panel B. Western blot analysis of induced samples using anti-HIS probe. Lanes (1), (2) and (3) are B. melitensis Omp31, BP26, and Omp16, respectively.
    Competent E Coli Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 2608 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher bl21 competent cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher bl21 a1 cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 A1 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bl21 bacterial cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 Bacterial Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher dh5 alpha cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Dh5 Alpha Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 167 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher oneshot bl21 cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Oneshot Bl21 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher bl21 de3 rosetta cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 De3 Rosetta Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bl21 startm de3 cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 Startm De3 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher bl21 startm cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 Startm Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher bl21 de3 plyss cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 De3 Plyss Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 135 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bl21 de3 gold cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 De3 Gold Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bl21 codonplus de3 cells
    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli <t>BL21</t> (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.
    Bl21 Codonplus De3 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    SDS-PAGE and western blot analysis of E. coli BL21-AI cells expressing B. melitensis Omp31, BP26, or 16. Panel A. SDS-PAGE analysis of E. coli BL21-AI cells harbouring pDESTOMPs of B. melitensis. (M) Marker lane, (1) uninduced, (2), (3) and (4) are Omp31, BP26, and Omp16, respectively. Panel B. Western blot analysis of induced samples using anti-HIS probe. Lanes (1), (2) and (3) are B. melitensis Omp31, BP26, and Omp16, respectively.

    Journal: Vaccine

    Article Title: Invasive Escherichia coli vaccines expressing Brucella melitensis outer membrane proteins 31 or 16 or periplasmic protein BP26 confer protection in mice challenged with B. melitensis

    doi: 10.1016/j.vaccine.2012.04.036

    Figure Lengend Snippet: SDS-PAGE and western blot analysis of E. coli BL21-AI cells expressing B. melitensis Omp31, BP26, or 16. Panel A. SDS-PAGE analysis of E. coli BL21-AI cells harbouring pDESTOMPs of B. melitensis. (M) Marker lane, (1) uninduced, (2), (3) and (4) are Omp31, BP26, and Omp16, respectively. Panel B. Western blot analysis of induced samples using anti-HIS probe. Lanes (1), (2) and (3) are B. melitensis Omp31, BP26, and Omp16, respectively.

    Article Snippet: Construction of inv E. coli vaccines used co-transformation of BL21-AI cells (Invitrogen) with pGB2Ωinv-hly and pDEST17/Omps.

    Techniques: SDS Page, Western Blot, Expressing, Marker

    Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli BL21 (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.

    Journal: Virulence

    Article Title: Rab18 binds to classical swine fever virus NS5A and mediates viral replication and assembly in swine umbilical vein endothelial cells

    doi: 10.1080/21505594.2020.1767356

    Figure Lengend Snippet: Rab18 binds to CSFV NS5A. (a) Co-immunoprecipitation (co-IP) assay. Cells were transfected with pcDNA-NS2-Myc or pcDNA-NS5A-Myc for 48 h. The transfected cells were lysed and immunoprecipitated, and western blot analysis was conducted using anti-Myc, anti-Rab18, and anti-β-actin. (b) GST-pulldown assay. GST or GST-Rab18 fusion proteins expressed in E. coli BL21 (DE3) were purified with glutathione agarose resin and incubated with the lysate of NS5A-Myc-expressing cells. Western blot analysis using anti-GST, anti-Myc, and anti-β-actin. (c) Exogenous NS5A-Myc binds to Rab18-Flag, S22N-Flag, and Q67 L-Flag in co-transfected cells. pcDNA-NS5A-Myc with pCDNA3.1-Rab18-Flag-, pCDNA3.1-S22N-Flag-, and pCDNA3.1-Q67L-Flag-transfected cells were lysed and immunoprecipitated. Then, western blot analysis was conducted using anti-Myc, anti-Flag, and anti-β-actin. pcDNA-NS2-Myc with pCDNA3.1-Rab18-Flag was used as a negative control. (d) Rab18 co-localization with CSFV NS5A protein. Cells were co-transfected with NS5A-GFP and Rab18-Red, NS5A-GFP and Q67L-Red, and NS5A-GFP and S22N-Red. Plasmids pEGFP-N1 and pCDNA-Red were co-transfected as a control. At 48 h after transfection, cells were fixed in 4% paraformaldehyde and stained with DAPI to label nuclei (blue). Scale bars, 10 μm.

    Article Snippet: GST-pulldown assaysThe plasmids pGEX-6p-1 and pGEX-GST-Rab18 were transformed into BL21 competent cells (Invitrogen, Carlsbad, CA, United States) to obtain GST and GST-Rab18 proteins, respectively.

    Techniques: Co-Immunoprecipitation Assay, Transfection, Immunoprecipitation, Western Blot, GST Pulldown Assay, Purification, Incubation, Expressing, Negative Control, Staining