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  • 94
    Thermo Fisher bl 21 de3 plyss one shot e coli
    Bl 21 De3 Plyss One Shot E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore bl 21 de3 plyss competent cells
    Bl 21 De3 Plyss Competent Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega bl 21 de3 plyss
    Expression of rROP1 in two different bacterial expression hosts, E. coli <t>BL21</t> (DE3) <t>pLysS</t> (lanes 1 and 2) and Rosetta (lanes 3 and 4). Induced (lanes 2 and 4) and uninduced bacteria (lanes 1 and 3) were analyzed on SDSPAGE. Densitometry analysis using Image J software showed rROP1band attributed to 1.5% and 0.5% of total protein in induced Rosetta (DE3) and BL21 (DE3) pLysS bacteria, respectively
    Bl 21 De3 Plyss, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bl21 de3 plyss
    Expression and purification of fusion protein. <t>1—pET16b–scFv-DAF/BL21</t> (DE3) <t>plyss</t> before induction; 2—pET16b–scFv-DAF induced by 1 mM IPTG for 4 h; 3 and 4—eluted peak of fusion protein from Hitrap chelating HP column; 5 and 6—pET16b–scFv-DAF refolded with the optimized urea gradient dialysis method. M, low molecular protein markers. The black arrow on lane 2 indicates the additional band at 61 kDa.
    Bl21 De3 Plyss, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 203 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene bl21 de3 plyss
    Overexpression of the recombinant T. pallidum Gpd and analysis of anti-Gpd immunoreactivity. (A) Coomassie blue-stained SDS-PAGE analysis of E. coli <t>BL21</t> (DE3) <t>pLysS</t> expressing either the Gpd-pET-3a construct (lane 1, crude lysate; lane 2, soluble fraction; lane 3, insoluble fraction) or the pET-3a vector alone (lane 4, insoluble fraction). (B) Immunoblot analysis of anti-Gpd immunoreactivity on purified inclusion bodies from E. coli BL21 (DE3) pLysS expressing the Gpd-pET-3a construct. Lanes: 1, anti-Gpd polyclonal antiserum, 2, E. coli -adsorbed anti-Gpd polyclonal antiserum. Each lane contains approximately 2 μg of total bacterial lysate and soluble or insoluble bacterial fractions, and molecular mass standards in kilodaltons are indicated at the left of each panel. In each panel, the recombinant T. pallidum Gpd is indicated by an arrow.
    Bl21 De3 Plyss, supplied by Stratagene, used in various techniques. Bioz Stars score: 91/100, based on 100 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore bl21 de3 plyss
    Overexpression of the recombinant T. pallidum Gpd and analysis of anti-Gpd immunoreactivity. (A) Coomassie blue-stained SDS-PAGE analysis of E. coli <t>BL21</t> (DE3) <t>pLysS</t> expressing either the Gpd-pET-3a construct (lane 1, crude lysate; lane 2, soluble fraction; lane 3, insoluble fraction) or the pET-3a vector alone (lane 4, insoluble fraction). (B) Immunoblot analysis of anti-Gpd immunoreactivity on purified inclusion bodies from E. coli BL21 (DE3) pLysS expressing the Gpd-pET-3a construct. Lanes: 1, anti-Gpd polyclonal antiserum, 2, E. coli -adsorbed anti-Gpd polyclonal antiserum. Each lane contains approximately 2 μg of total bacterial lysate and soluble or insoluble bacterial fractions, and molecular mass standards in kilodaltons are indicated at the left of each panel. In each panel, the recombinant T. pallidum Gpd is indicated by an arrow.
    Bl21 De3 Plyss, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 622 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies bl21 de3 plyss
    Enzymatic activity assays using glucanase- and xylanase-IFP fusion proteins expressed in E. coli . LIC-pDEST-LC1-/LC2 vectors encoding 6xHis-IFP-TEV-endo-β-1,4-glucanase/-xylanase and endo-β-1,4-glucanase-/xylanase-TEV-IFP-6xHis fusion proteins were transformed into the E. coli strains <t>BL21</t> (DE3) <t>pLysS</t> (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’) and Rosetta (DE3) pRARE (‘Rosetta’). Enzymatic activity was tested by Congo Red staining and destaining with 1 M NaCl on carboxymethylcellulose- (left panel) or xylan- (right panel) containing agar plates after transferring 2 µL of the respective expression strains and over-night incubation at 37°C. Glucanase activity leads to the formation of a white halo around the colonies, whereas xylanase activity leads to the formation of a black halo [31] . E. coli cells expressing IFP-6xHis fusion protein were used as negative control, and cell-free supernatant of Pichia pastoris expression cultures containing secreted endo-β-1,4-glucanase-myc-6xHis or endo-β-1,4-xylanase-myc-6xHis fusion proteins were used as positive controls (P).
    Bl21 De3 Plyss, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA bl21 de3 plyss
    Enzymatic activity assays using glucanase- and xylanase-IFP fusion proteins expressed in E. coli . LIC-pDEST-LC1-/LC2 vectors encoding 6xHis-IFP-TEV-endo-β-1,4-glucanase/-xylanase and endo-β-1,4-glucanase-/xylanase-TEV-IFP-6xHis fusion proteins were transformed into the E. coli strains <t>BL21</t> (DE3) <t>pLysS</t> (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’) and Rosetta (DE3) pRARE (‘Rosetta’). Enzymatic activity was tested by Congo Red staining and destaining with 1 M NaCl on carboxymethylcellulose- (left panel) or xylan- (right panel) containing agar plates after transferring 2 µL of the respective expression strains and over-night incubation at 37°C. Glucanase activity leads to the formation of a white halo around the colonies, whereas xylanase activity leads to the formation of a black halo [31] . E. coli cells expressing IFP-6xHis fusion protein were used as negative control, and cell-free supernatant of Pichia pastoris expression cultures containing secreted endo-β-1,4-glucanase-myc-6xHis or endo-β-1,4-xylanase-myc-6xHis fusion proteins were used as positive controls (P).
    Bl21 De3 Plyss, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 91/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene bl21 gold de3 plyss
    Enzymatic activity assays using glucanase- and xylanase-IFP fusion proteins expressed in E. coli . LIC-pDEST-LC1-/LC2 vectors encoding 6xHis-IFP-TEV-endo-β-1,4-glucanase/-xylanase and endo-β-1,4-glucanase-/xylanase-TEV-IFP-6xHis fusion proteins were transformed into the E. coli strains <t>BL21</t> (DE3) <t>pLysS</t> (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’) and Rosetta (DE3) pRARE (‘Rosetta’). Enzymatic activity was tested by Congo Red staining and destaining with 1 M NaCl on carboxymethylcellulose- (left panel) or xylan- (right panel) containing agar plates after transferring 2 µL of the respective expression strains and over-night incubation at 37°C. Glucanase activity leads to the formation of a white halo around the colonies, whereas xylanase activity leads to the formation of a black halo [31] . E. coli cells expressing IFP-6xHis fusion protein were used as negative control, and cell-free supernatant of Pichia pastoris expression cultures containing secreted endo-β-1,4-glucanase-myc-6xHis or endo-β-1,4-xylanase-myc-6xHis fusion proteins were used as positive controls (P).
    Bl21 Gold De3 Plyss, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA competent bl21 de3 plyss
    Enzymatic activity assays using glucanase- and xylanase-IFP fusion proteins expressed in E. coli . LIC-pDEST-LC1-/LC2 vectors encoding 6xHis-IFP-TEV-endo-β-1,4-glucanase/-xylanase and endo-β-1,4-glucanase-/xylanase-TEV-IFP-6xHis fusion proteins were transformed into the E. coli strains <t>BL21</t> (DE3) <t>pLysS</t> (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’) and Rosetta (DE3) pRARE (‘Rosetta’). Enzymatic activity was tested by Congo Red staining and destaining with 1 M NaCl on carboxymethylcellulose- (left panel) or xylan- (right panel) containing agar plates after transferring 2 µL of the respective expression strains and over-night incubation at 37°C. Glucanase activity leads to the formation of a white halo around the colonies, whereas xylanase activity leads to the formation of a black halo [31] . E. coli cells expressing IFP-6xHis fusion protein were used as negative control, and cell-free supernatant of Pichia pastoris expression cultures containing secreted endo-β-1,4-glucanase-myc-6xHis or endo-β-1,4-xylanase-myc-6xHis fusion proteins were used as positive controls (P).
    Competent Bl21 De3 Plyss, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Expression of rROP1 in two different bacterial expression hosts, E. coli BL21 (DE3) pLysS (lanes 1 and 2) and Rosetta (lanes 3 and 4). Induced (lanes 2 and 4) and uninduced bacteria (lanes 1 and 3) were analyzed on SDSPAGE. Densitometry analysis using Image J software showed rROP1band attributed to 1.5% and 0.5% of total protein in induced Rosetta (DE3) and BL21 (DE3) pLysS bacteria, respectively

    Journal: Avicenna Journal of Medical Biotechnology

    Article Title: Expression and Purification of Recombinant ROP1 of Toxoplasma gondii in Bacteria

    doi:

    Figure Lengend Snippet: Expression of rROP1 in two different bacterial expression hosts, E. coli BL21 (DE3) pLysS (lanes 1 and 2) and Rosetta (lanes 3 and 4). Induced (lanes 2 and 4) and uninduced bacteria (lanes 1 and 3) were analyzed on SDSPAGE. Densitometry analysis using Image J software showed rROP1band attributed to 1.5% and 0.5% of total protein in induced Rosetta (DE3) and BL21 (DE3) pLysS bacteria, respectively

    Article Snippet: E. coli Top 10F’ (Invitrogen, Carlsbad, CA, USA), E. coli BL21 (DE3) PlysS and Rosetta (DE3) (Promega, Madison, WI, USA) were used for cloning and expression of recombinant antigen, respectively.

    Techniques: Expressing, Software

    Expression and purification of fusion protein. 1—pET16b–scFv-DAF/BL21 (DE3) plyss before induction; 2—pET16b–scFv-DAF induced by 1 mM IPTG for 4 h; 3 and 4—eluted peak of fusion protein from Hitrap chelating HP column; 5 and 6—pET16b–scFv-DAF refolded with the optimized urea gradient dialysis method. M, low molecular protein markers. The black arrow on lane 2 indicates the additional band at 61 kDa.

    Journal: Muscle & nerve

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS

    doi: 10.1002/mus.23247

    Figure Lengend Snippet: Expression and purification of fusion protein. 1—pET16b–scFv-DAF/BL21 (DE3) plyss before induction; 2—pET16b–scFv-DAF induced by 1 mM IPTG for 4 h; 3 and 4—eluted peak of fusion protein from Hitrap chelating HP column; 5 and 6—pET16b–scFv-DAF refolded with the optimized urea gradient dialysis method. M, low molecular protein markers. The black arrow on lane 2 indicates the additional band at 61 kDa.

    Article Snippet: BL21 (DE3) pLyss (Invitrogen) bacteria transformed with the reconstructed plasmid pET16b were grown at 37°C in 2× YT medium containing 100 μ g/ml ampicillin and 50 μ g/ml chloramphenicol.

    Techniques: Expressing, Purification

    Overexpression of the recombinant T. pallidum Gpd and analysis of anti-Gpd immunoreactivity. (A) Coomassie blue-stained SDS-PAGE analysis of E. coli BL21 (DE3) pLysS expressing either the Gpd-pET-3a construct (lane 1, crude lysate; lane 2, soluble fraction; lane 3, insoluble fraction) or the pET-3a vector alone (lane 4, insoluble fraction). (B) Immunoblot analysis of anti-Gpd immunoreactivity on purified inclusion bodies from E. coli BL21 (DE3) pLysS expressing the Gpd-pET-3a construct. Lanes: 1, anti-Gpd polyclonal antiserum, 2, E. coli -adsorbed anti-Gpd polyclonal antiserum. Each lane contains approximately 2 μg of total bacterial lysate and soluble or insoluble bacterial fractions, and molecular mass standards in kilodaltons are indicated at the left of each panel. In each panel, the recombinant T. pallidum Gpd is indicated by an arrow.

    Journal: Infection and Immunity

    Article Title: Function and Protective Capacity of Treponema pallidum subsp. pallidum Glycerophosphodiester Phosphodiesterase

    doi:

    Figure Lengend Snippet: Overexpression of the recombinant T. pallidum Gpd and analysis of anti-Gpd immunoreactivity. (A) Coomassie blue-stained SDS-PAGE analysis of E. coli BL21 (DE3) pLysS expressing either the Gpd-pET-3a construct (lane 1, crude lysate; lane 2, soluble fraction; lane 3, insoluble fraction) or the pET-3a vector alone (lane 4, insoluble fraction). (B) Immunoblot analysis of anti-Gpd immunoreactivity on purified inclusion bodies from E. coli BL21 (DE3) pLysS expressing the Gpd-pET-3a construct. Lanes: 1, anti-Gpd polyclonal antiserum, 2, E. coli -adsorbed anti-Gpd polyclonal antiserum. Each lane contains approximately 2 μg of total bacterial lysate and soluble or insoluble bacterial fractions, and molecular mass standards in kilodaltons are indicated at the left of each panel. In each panel, the recombinant T. pallidum Gpd is indicated by an arrow.

    Article Snippet: E. coli XL-1 Blue and BL21 (DE3) pLysS were obtained from Stratagene (La Jolla, Calif.).

    Techniques: Over Expression, Recombinant, Staining, SDS Page, Expressing, Positron Emission Tomography, Construct, Plasmid Preparation, Purification

    Immunoblot analyses of the Ig-binding capability of recombinant T. pallidum Gpd. Shown are inclusion bodies purified from E. coli BL21 (DE3) pLysS transformed with either the pET-3a-Gpd construct (lanes 1, 3, 5, and 7) or with the pET-3a vector alone (lanes 2, 4, 6, and 8). Molecular mass standards in kilodaltons are indicated on the left of each panel. (A) Specificity of recombinant T. pallidum Gpd for binding various Ig classes. Lanes: 1 and 2, human IgA and goat F(ab′) 2 anti-human IgA (alpha chain specific); 3 and 4, human IgD and goat F(ab′) 2 anti-human IgD (delta chain specific); 5 and 6, human IgG and goat F(ab′) 2 anti-human IgG (gamma chain specific); 7 and 8, human IgM and goat F(ab′) 2 anti-human IgM (mu chain specific). (B) Investigation of the binding specificity of recombinant T. pallidum Gpd for human IgG. In all cases the secondary antibody used was goat F(ab′) 2 anti-human IgG (gamma chain specific). Lanes: 1 and 2, intact human IgG; 3 and 4, human IgG Fc fragment; 5 and 6, human IgG Fab fragment; 7 and 8, no primary Ig.

    Journal: Infection and Immunity

    Article Title: Function and Protective Capacity of Treponema pallidum subsp. pallidum Glycerophosphodiester Phosphodiesterase

    doi:

    Figure Lengend Snippet: Immunoblot analyses of the Ig-binding capability of recombinant T. pallidum Gpd. Shown are inclusion bodies purified from E. coli BL21 (DE3) pLysS transformed with either the pET-3a-Gpd construct (lanes 1, 3, 5, and 7) or with the pET-3a vector alone (lanes 2, 4, 6, and 8). Molecular mass standards in kilodaltons are indicated on the left of each panel. (A) Specificity of recombinant T. pallidum Gpd for binding various Ig classes. Lanes: 1 and 2, human IgA and goat F(ab′) 2 anti-human IgA (alpha chain specific); 3 and 4, human IgD and goat F(ab′) 2 anti-human IgD (delta chain specific); 5 and 6, human IgG and goat F(ab′) 2 anti-human IgG (gamma chain specific); 7 and 8, human IgM and goat F(ab′) 2 anti-human IgM (mu chain specific). (B) Investigation of the binding specificity of recombinant T. pallidum Gpd for human IgG. In all cases the secondary antibody used was goat F(ab′) 2 anti-human IgG (gamma chain specific). Lanes: 1 and 2, intact human IgG; 3 and 4, human IgG Fc fragment; 5 and 6, human IgG Fab fragment; 7 and 8, no primary Ig.

    Article Snippet: E. coli XL-1 Blue and BL21 (DE3) pLysS were obtained from Stratagene (La Jolla, Calif.).

    Techniques: Binding Assay, Recombinant, Purification, Transformation Assay, Positron Emission Tomography, Construct, Plasmid Preparation

    Enzymatic activity assays using glucanase- and xylanase-IFP fusion proteins expressed in E. coli . LIC-pDEST-LC1-/LC2 vectors encoding 6xHis-IFP-TEV-endo-β-1,4-glucanase/-xylanase and endo-β-1,4-glucanase-/xylanase-TEV-IFP-6xHis fusion proteins were transformed into the E. coli strains BL21 (DE3) pLysS (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’) and Rosetta (DE3) pRARE (‘Rosetta’). Enzymatic activity was tested by Congo Red staining and destaining with 1 M NaCl on carboxymethylcellulose- (left panel) or xylan- (right panel) containing agar plates after transferring 2 µL of the respective expression strains and over-night incubation at 37°C. Glucanase activity leads to the formation of a white halo around the colonies, whereas xylanase activity leads to the formation of a black halo [31] . E. coli cells expressing IFP-6xHis fusion protein were used as negative control, and cell-free supernatant of Pichia pastoris expression cultures containing secreted endo-β-1,4-glucanase-myc-6xHis or endo-β-1,4-xylanase-myc-6xHis fusion proteins were used as positive controls (P).

    Journal: PLoS ONE

    Article Title: High-Throughput Protein Expression Using a Combination of Ligation-Independent Cloning (LIC) and Infrared Fluorescent Protein (IFP) Detection

    doi: 10.1371/journal.pone.0018900

    Figure Lengend Snippet: Enzymatic activity assays using glucanase- and xylanase-IFP fusion proteins expressed in E. coli . LIC-pDEST-LC1-/LC2 vectors encoding 6xHis-IFP-TEV-endo-β-1,4-glucanase/-xylanase and endo-β-1,4-glucanase-/xylanase-TEV-IFP-6xHis fusion proteins were transformed into the E. coli strains BL21 (DE3) pLysS (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’) and Rosetta (DE3) pRARE (‘Rosetta’). Enzymatic activity was tested by Congo Red staining and destaining with 1 M NaCl on carboxymethylcellulose- (left panel) or xylan- (right panel) containing agar plates after transferring 2 µL of the respective expression strains and over-night incubation at 37°C. Glucanase activity leads to the formation of a white halo around the colonies, whereas xylanase activity leads to the formation of a black halo [31] . E. coli cells expressing IFP-6xHis fusion protein were used as negative control, and cell-free supernatant of Pichia pastoris expression cultures containing secreted endo-β-1,4-glucanase-myc-6xHis or endo-β-1,4-xylanase-myc-6xHis fusion proteins were used as positive controls (P).

    Article Snippet: Protein expression for the purification of GST (empty pDEST15 vector) or GST-GRF fusion proteins (pDEST15-GRF1/2/3/4/5/6 vectors) was carried out in 100 mL culture volume in BL21 (DE3) pLysS (Agilent Technologies) cells (30°C, 1 mM IPTG, 4 h), followed by sonication of cells in 10 mL lysis buffer as described above.

    Techniques: Activity Assay, Transformation Assay, Staining, Transferring, Expressing, Incubation, Negative Control

    Infrared analysis of in vitro and in vivo expressed IFP fusion proteins. Infrared scanning of all samples was performed in microtiter plates using the Odyssey Infrared Imaging System from LI-COR Biosciences. ( A ) In vitro transcription/translation products were analysed by infrared scanning using the whole reaction mixtures. 6xHis-GFP and IFP-6xHis fusion protein-expressing samples were used as negative (-) and positive (+) controls, respectively. ( B ) In-cell detection of IFP fusion protein (6xHis-IFP-TEV-ANAC042 shown as an example) in two randomly selected clones each from the E. coli strains (BL21 (DE3) pLysS (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’), and Rosetta (DE3) pRARE (‘Rosetta’). 6xHis-GFP and IFP-6xHis fusion protein-expressing cells were used as negative (-) and positive (+) controls, respectively. ( C ), ( D ) and ( E ) In-cell detection of IFP fusion protein (SAM1-TEV-IFP-6xHis shown as an example) in randomly selected K. lactis , P. pastoris and L. tarentolae clones. Cell lines not expressing IPF or expressing IFP-6xHis fusion protein were used as negative (-) and positive (+) controls, respectively. No positive control was available for expression in K. lactis . Note that strong infrared signal appears white in the digital images.

    Journal: PLoS ONE

    Article Title: High-Throughput Protein Expression Using a Combination of Ligation-Independent Cloning (LIC) and Infrared Fluorescent Protein (IFP) Detection

    doi: 10.1371/journal.pone.0018900

    Figure Lengend Snippet: Infrared analysis of in vitro and in vivo expressed IFP fusion proteins. Infrared scanning of all samples was performed in microtiter plates using the Odyssey Infrared Imaging System from LI-COR Biosciences. ( A ) In vitro transcription/translation products were analysed by infrared scanning using the whole reaction mixtures. 6xHis-GFP and IFP-6xHis fusion protein-expressing samples were used as negative (-) and positive (+) controls, respectively. ( B ) In-cell detection of IFP fusion protein (6xHis-IFP-TEV-ANAC042 shown as an example) in two randomly selected clones each from the E. coli strains (BL21 (DE3) pLysS (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’), and Rosetta (DE3) pRARE (‘Rosetta’). 6xHis-GFP and IFP-6xHis fusion protein-expressing cells were used as negative (-) and positive (+) controls, respectively. ( C ), ( D ) and ( E ) In-cell detection of IFP fusion protein (SAM1-TEV-IFP-6xHis shown as an example) in randomly selected K. lactis , P. pastoris and L. tarentolae clones. Cell lines not expressing IPF or expressing IFP-6xHis fusion protein were used as negative (-) and positive (+) controls, respectively. No positive control was available for expression in K. lactis . Note that strong infrared signal appears white in the digital images.

    Article Snippet: Protein expression for the purification of GST (empty pDEST15 vector) or GST-GRF fusion proteins (pDEST15-GRF1/2/3/4/5/6 vectors) was carried out in 100 mL culture volume in BL21 (DE3) pLysS (Agilent Technologies) cells (30°C, 1 mM IPTG, 4 h), followed by sonication of cells in 10 mL lysis buffer as described above.

    Techniques: In Vitro, In Vivo, Imaging, Expressing, Clone Assay, Positive Control

    Expression of IFP fusion proteins in E. coli . Protein extracts obtained from IFP fusion protein-expressing E. coli strains BL21 (DE3) pLysS (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’), and Rosetta (DE3) pRARE (‘Rosetta’) were separated by SDS-PAGE and analysed by in-gel infrared imaging at 700 nm to detect IFP moieties (upper panel), followed by western transfer and immunological detection at 800 nm (using monoclonal mouse antibody directed against the 6xHis epitope; lower panel). IFP-6xHis fusion protein-expressing cells were used as positive control. Supernatant (S) and pellet (P) fractions of disrupted cells were analyzed after ultracentrifugation. M, molecular mass marker (kDa). Arrows indicate expected proteins.

    Journal: PLoS ONE

    Article Title: High-Throughput Protein Expression Using a Combination of Ligation-Independent Cloning (LIC) and Infrared Fluorescent Protein (IFP) Detection

    doi: 10.1371/journal.pone.0018900

    Figure Lengend Snippet: Expression of IFP fusion proteins in E. coli . Protein extracts obtained from IFP fusion protein-expressing E. coli strains BL21 (DE3) pLysS (‘pLysS’), BL21 Star (DE3) pRARE (‘Star’), BL21 (DE3) CodonPlus-RIL (‘Codon’), and Rosetta (DE3) pRARE (‘Rosetta’) were separated by SDS-PAGE and analysed by in-gel infrared imaging at 700 nm to detect IFP moieties (upper panel), followed by western transfer and immunological detection at 800 nm (using monoclonal mouse antibody directed against the 6xHis epitope; lower panel). IFP-6xHis fusion protein-expressing cells were used as positive control. Supernatant (S) and pellet (P) fractions of disrupted cells were analyzed after ultracentrifugation. M, molecular mass marker (kDa). Arrows indicate expected proteins.

    Article Snippet: Protein expression for the purification of GST (empty pDEST15 vector) or GST-GRF fusion proteins (pDEST15-GRF1/2/3/4/5/6 vectors) was carried out in 100 mL culture volume in BL21 (DE3) pLysS (Agilent Technologies) cells (30°C, 1 mM IPTG, 4 h), followed by sonication of cells in 10 mL lysis buffer as described above.

    Techniques: Expressing, SDS Page, Imaging, Western Blot, Positive Control, Marker