bl21 de3 Millipore Search Results


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  • 95
    Millipore e coli bl21
    RP-HPLC separation of the heated cellular extract. Cellular extract of E. coli <t>BL21</t> [DE3] (pMA24) were treated as described in Section 2.4 . Chromatogram was monitored at (A) λ = 220 nm and (B) λ = 280 nm. Peaks were collected and individually analyzed by MALDI-TOF MS.
    E Coli Bl21, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 9228 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore bl21 de3 competent cells
    RP-HPLC separation of the heated cellular extract. Cellular extract of E. coli <t>BL21</t> [DE3] (pMA24) were treated as described in Section 2.4 . Chromatogram was monitored at (A) λ = 220 nm and (B) λ = 280 nm. Peaks were collected and individually analyzed by MALDI-TOF MS.
    Bl21 De3 Competent Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 208 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore bl21 de3 plyss
    Expression of UspF protein by an IPTG‐inducible E. coli <t>BL21</t> (DE3) <t>pLyS</t> strain and purification of soluble fractions containing UspF protein. A – T0 total protein extract before induction; T2, total protein extract after induction (3 h); T3, total protein extract after induction (16–18 h). The arrow indicates position of the UspF protein (18.4 kDa). B – Purification of soluble fractions or UspF protein by affinity chromatography using a nickel‐containing resin. 1–Flow through; 2–30 m M , 3–50 m M , 4–100 m M ; 5–200 m M of imidazole.
    Bl21 De3 Plyss, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 557 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Millipore e coli bl21 de3 plyss competent cells
    Expression of UspF protein by an IPTG‐inducible E. coli <t>BL21</t> (DE3) <t>pLyS</t> strain and purification of soluble fractions containing UspF protein. A – T0 total protein extract before induction; T2, total protein extract after induction (3 h); T3, total protein extract after induction (16–18 h). The arrow indicates position of the UspF protein (18.4 kDa). B – Purification of soluble fractions or UspF protein by affinity chromatography using a nickel‐containing resin. 1–Flow through; 2–30 m M , 3–50 m M , 4–100 m M ; 5–200 m M of imidazole.
    E Coli Bl21 De3 Plyss Competent Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 82 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Millipore escherichiacoli bl21
    Expression of UspF protein by an IPTG‐inducible E. coli <t>BL21</t> (DE3) <t>pLyS</t> strain and purification of soluble fractions containing UspF protein. A – T0 total protein extract before induction; T2, total protein extract after induction (3 h); T3, total protein extract after induction (16–18 h). The arrow indicates position of the UspF protein (18.4 kDa). B – Purification of soluble fractions or UspF protein by affinity chromatography using a nickel‐containing resin. 1–Flow through; 2–30 m M , 3–50 m M , 4–100 m M ; 5–200 m M of imidazole.
    Escherichiacoli Bl21, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Millipore bl21 de3 ril
    Expression of UspF protein by an IPTG‐inducible E. coli <t>BL21</t> (DE3) <t>pLyS</t> strain and purification of soluble fractions containing UspF protein. A – T0 total protein extract before induction; T2, total protein extract after induction (3 h); T3, total protein extract after induction (16–18 h). The arrow indicates position of the UspF protein (18.4 kDa). B – Purification of soluble fractions or UspF protein by affinity chromatography using a nickel‐containing resin. 1–Flow through; 2–30 m M , 3–50 m M , 4–100 m M ; 5–200 m M of imidazole.
    Bl21 De3 Ril, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Merck KGaA bl21
    Expression of UspF protein by an IPTG‐inducible E. coli <t>BL21</t> (DE3) <t>pLyS</t> strain and purification of soluble fractions containing UspF protein. A – T0 total protein extract before induction; T2, total protein extract after induction (3 h); T3, total protein extract after induction (16–18 h). The arrow indicates position of the UspF protein (18.4 kDa). B – Purification of soluble fractions or UspF protein by affinity chromatography using a nickel‐containing resin. 1–Flow through; 2–30 m M , 3–50 m M , 4–100 m M ; 5–200 m M of imidazole.
    Bl21, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 94/100, based on 125 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Millipore bl21 tuner de3
    A fly SUMO-ome: characterization and validation. A) Scheme for identifying Drosophila embryonic SUMO conjugates. SUMO conjugates were isolated by tandem affinity purification from transgenic fly embryos expressing (His) 6 -FLAG-SUMO. The initial purification step (Ni-NTA chromatography) was performed under denaturing conditions. To maximize the chance of detecting low abundance proteins in the complex protein mixture, the affinity-purified proteins were separated by SDS-PAGE, and the lane was cut into 20 evenly divided gel slices. Tryptic peptides extracted from each gel slice were analyzed by LC-MS/MS. B) A bacterial sumoylation assay. The Q SUMO vector, which encodes the mature form of SUMO (SUMO GG ) along with SAE1, SAE2, and Ubc9 expressed from separate T7/lac promoters, was used in combination with a vector expressing a GST-tagged candidate substrate. As a negative control, Q ΔGG , which expresses a conjugation defective form of SUMO (SUMO ΔGG ), was used in place of Q SUMO . C) Bacterial sumoylation assays were used to validate proteins identified in the proteomic screen as sumoylation substrates. GST-tagged candidate SUMO conjugates were expressed in <t>BL21</t> cells co-transformed with Q SUMO or Q ΔGG vectors, purified using glutathione beads, and immunoblotted using antibodies against GST, SUMO, or poly-His (to detect 6xHis-tagged SUMO). GST by itself was not sumoylated in this assay. Black arrows point to the bands representing sumoylated proteins, and open arrow points to a non-specific reacting band. D) The eIF4E protein was purified from Drosophila S2 cells stably expressing FLAG-(His) 6 - tagged eIF4E using Ni-NTA beads under denaturing conditions. The resulting proteins were probed with anti-FLAG antibody in a Western blot. The cells were treated with SUMO or control YFP dsRNA for 3 days prior to cell lysis. In the control sample, the bands representing the sumoylated species (black arrows) have intensities that are 8.1% (top) and 12.9% (bottom) of the intensity of the band representing unmodified eIF4E (∼40 kDa), whereas in the SUMO knockdown sample, they are reduced to 1.8% (top) and 3.5% (bottom). Quantitation was performed using Quantity One 4.3.0 (BioRad).
    Bl21 Tuner De3, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Millipore rosetta bl21
    A fly SUMO-ome: characterization and validation. A) Scheme for identifying Drosophila embryonic SUMO conjugates. SUMO conjugates were isolated by tandem affinity purification from transgenic fly embryos expressing (His) 6 -FLAG-SUMO. The initial purification step (Ni-NTA chromatography) was performed under denaturing conditions. To maximize the chance of detecting low abundance proteins in the complex protein mixture, the affinity-purified proteins were separated by SDS-PAGE, and the lane was cut into 20 evenly divided gel slices. Tryptic peptides extracted from each gel slice were analyzed by LC-MS/MS. B) A bacterial sumoylation assay. The Q SUMO vector, which encodes the mature form of SUMO (SUMO GG ) along with SAE1, SAE2, and Ubc9 expressed from separate T7/lac promoters, was used in combination with a vector expressing a GST-tagged candidate substrate. As a negative control, Q ΔGG , which expresses a conjugation defective form of SUMO (SUMO ΔGG ), was used in place of Q SUMO . C) Bacterial sumoylation assays were used to validate proteins identified in the proteomic screen as sumoylation substrates. GST-tagged candidate SUMO conjugates were expressed in <t>BL21</t> cells co-transformed with Q SUMO or Q ΔGG vectors, purified using glutathione beads, and immunoblotted using antibodies against GST, SUMO, or poly-His (to detect 6xHis-tagged SUMO). GST by itself was not sumoylated in this assay. Black arrows point to the bands representing sumoylated proteins, and open arrow points to a non-specific reacting band. D) The eIF4E protein was purified from Drosophila S2 cells stably expressing FLAG-(His) 6 - tagged eIF4E using Ni-NTA beads under denaturing conditions. The resulting proteins were probed with anti-FLAG antibody in a Western blot. The cells were treated with SUMO or control YFP dsRNA for 3 days prior to cell lysis. In the control sample, the bands representing the sumoylated species (black arrows) have intensities that are 8.1% (top) and 12.9% (bottom) of the intensity of the band representing unmodified eIF4E (∼40 kDa), whereas in the SUMO knockdown sample, they are reduced to 1.8% (top) and 3.5% (bottom). Quantitation was performed using Quantity One 4.3.0 (BioRad).
    Rosetta Bl21, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore bl21 de3 r3 rosetta
    A fly SUMO-ome: characterization and validation. A) Scheme for identifying Drosophila embryonic SUMO conjugates. SUMO conjugates were isolated by tandem affinity purification from transgenic fly embryos expressing (His) 6 -FLAG-SUMO. The initial purification step (Ni-NTA chromatography) was performed under denaturing conditions. To maximize the chance of detecting low abundance proteins in the complex protein mixture, the affinity-purified proteins were separated by SDS-PAGE, and the lane was cut into 20 evenly divided gel slices. Tryptic peptides extracted from each gel slice were analyzed by LC-MS/MS. B) A bacterial sumoylation assay. The Q SUMO vector, which encodes the mature form of SUMO (SUMO GG ) along with SAE1, SAE2, and Ubc9 expressed from separate T7/lac promoters, was used in combination with a vector expressing a GST-tagged candidate substrate. As a negative control, Q ΔGG , which expresses a conjugation defective form of SUMO (SUMO ΔGG ), was used in place of Q SUMO . C) Bacterial sumoylation assays were used to validate proteins identified in the proteomic screen as sumoylation substrates. GST-tagged candidate SUMO conjugates were expressed in <t>BL21</t> cells co-transformed with Q SUMO or Q ΔGG vectors, purified using glutathione beads, and immunoblotted using antibodies against GST, SUMO, or poly-His (to detect 6xHis-tagged SUMO). GST by itself was not sumoylated in this assay. Black arrows point to the bands representing sumoylated proteins, and open arrow points to a non-specific reacting band. D) The eIF4E protein was purified from Drosophila S2 cells stably expressing FLAG-(His) 6 - tagged eIF4E using Ni-NTA beads under denaturing conditions. The resulting proteins were probed with anti-FLAG antibody in a Western blot. The cells were treated with SUMO or control YFP dsRNA for 3 days prior to cell lysis. In the control sample, the bands representing the sumoylated species (black arrows) have intensities that are 8.1% (top) and 12.9% (bottom) of the intensity of the band representing unmodified eIF4E (∼40 kDa), whereas in the SUMO knockdown sample, they are reduced to 1.8% (top) and 3.5% (bottom). Quantitation was performed using Quantity One 4.3.0 (BioRad).
    Bl21 De3 R3 Rosetta, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Millipore bacterial bl21 de3
    A fly SUMO-ome: characterization and validation. A) Scheme for identifying Drosophila embryonic SUMO conjugates. SUMO conjugates were isolated by tandem affinity purification from transgenic fly embryos expressing (His) 6 -FLAG-SUMO. The initial purification step (Ni-NTA chromatography) was performed under denaturing conditions. To maximize the chance of detecting low abundance proteins in the complex protein mixture, the affinity-purified proteins were separated by SDS-PAGE, and the lane was cut into 20 evenly divided gel slices. Tryptic peptides extracted from each gel slice were analyzed by LC-MS/MS. B) A bacterial sumoylation assay. The Q SUMO vector, which encodes the mature form of SUMO (SUMO GG ) along with SAE1, SAE2, and Ubc9 expressed from separate T7/lac promoters, was used in combination with a vector expressing a GST-tagged candidate substrate. As a negative control, Q ΔGG , which expresses a conjugation defective form of SUMO (SUMO ΔGG ), was used in place of Q SUMO . C) Bacterial sumoylation assays were used to validate proteins identified in the proteomic screen as sumoylation substrates. GST-tagged candidate SUMO conjugates were expressed in <t>BL21</t> cells co-transformed with Q SUMO or Q ΔGG vectors, purified using glutathione beads, and immunoblotted using antibodies against GST, SUMO, or poly-His (to detect 6xHis-tagged SUMO). GST by itself was not sumoylated in this assay. Black arrows point to the bands representing sumoylated proteins, and open arrow points to a non-specific reacting band. D) The eIF4E protein was purified from Drosophila S2 cells stably expressing FLAG-(His) 6 - tagged eIF4E using Ni-NTA beads under denaturing conditions. The resulting proteins were probed with anti-FLAG antibody in a Western blot. The cells were treated with SUMO or control YFP dsRNA for 3 days prior to cell lysis. In the control sample, the bands representing the sumoylated species (black arrows) have intensities that are 8.1% (top) and 12.9% (bottom) of the intensity of the band representing unmodified eIF4E (∼40 kDa), whereas in the SUMO knockdown sample, they are reduced to 1.8% (top) and 3.5% (bottom). Quantitation was performed using Quantity One 4.3.0 (BioRad).
    Bacterial Bl21 De3, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore bl21 de3 rosetta
    Optimization of conditions for the overexpression of ELIC 1a . A cartoon representation of ELIC highlighting the membrane spanning pore forming domain (red helices), the extracellular domain (yellow B-sheets), and the boundaries of the cell membrane in relation to the channel structure. 1b . Upper panel, a western blot analysis of the expression levels of the ELIC channel made in the E. coli strains: C41, C43, <t>BL21</t> Gold, BL21 Codon (+) and BL21 <t>Rosetta</t> in the following growth media: Terrific Broth (TB, yellow), Auto-Induction medium (AI, blue) and LB medium (red). Lower panel, normalized ELIC band density from the western blot (Penta-histidine antibody) were plotted against E. coli strains grouped by growth media. 1c. The effect of the cell culture volume on ELIC expression levels and final culture biomass was studied for the three most promising E. coli strains: BL21 Gold, BL21 Codon (+) and BL21 Rosetta. 1d. The effect of Zn 2+ and Mg 2+ as channel negative modifiers or the competitive antagonist ACh on ELIC expression levels is represented as a bar plot.
    Bl21 De3 Rosetta, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Millipore bl21 de3 trxb
    Optimization of conditions for the overexpression of ELIC 1a . A cartoon representation of ELIC highlighting the membrane spanning pore forming domain (red helices), the extracellular domain (yellow B-sheets), and the boundaries of the cell membrane in relation to the channel structure. 1b . Upper panel, a western blot analysis of the expression levels of the ELIC channel made in the E. coli strains: C41, C43, <t>BL21</t> Gold, BL21 Codon (+) and BL21 <t>Rosetta</t> in the following growth media: Terrific Broth (TB, yellow), Auto-Induction medium (AI, blue) and LB medium (red). Lower panel, normalized ELIC band density from the western blot (Penta-histidine antibody) were plotted against E. coli strains grouped by growth media. 1c. The effect of the cell culture volume on ELIC expression levels and final culture biomass was studied for the three most promising E. coli strains: BL21 Gold, BL21 Codon (+) and BL21 Rosetta. 1d. The effect of Zn 2+ and Mg 2+ as channel negative modifiers or the competitive antagonist ACh on ELIC expression levels is represented as a bar plot.
    Bl21 De3 Trxb, supplied by Millipore, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore rosetta2
    Optimization of conditions for the overexpression of ELIC 1a . A cartoon representation of ELIC highlighting the membrane spanning pore forming domain (red helices), the extracellular domain (yellow B-sheets), and the boundaries of the cell membrane in relation to the channel structure. 1b . Upper panel, a western blot analysis of the expression levels of the ELIC channel made in the E. coli strains: C41, C43, <t>BL21</t> Gold, BL21 Codon (+) and BL21 <t>Rosetta</t> in the following growth media: Terrific Broth (TB, yellow), Auto-Induction medium (AI, blue) and LB medium (red). Lower panel, normalized ELIC band density from the western blot (Penta-histidine antibody) were plotted against E. coli strains grouped by growth media. 1c. The effect of the cell culture volume on ELIC expression levels and final culture biomass was studied for the three most promising E. coli strains: BL21 Gold, BL21 Codon (+) and BL21 Rosetta. 1d. The effect of Zn 2+ and Mg 2+ as channel negative modifiers or the competitive antagonist ACh on ELIC expression levels is represented as a bar plot.
    Rosetta2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1224 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Millipore induction control e bl21 de3
    Optimization of conditions for the overexpression of ELIC 1a . A cartoon representation of ELIC highlighting the membrane spanning pore forming domain (red helices), the extracellular domain (yellow B-sheets), and the boundaries of the cell membrane in relation to the channel structure. 1b . Upper panel, a western blot analysis of the expression levels of the ELIC channel made in the E. coli strains: C41, C43, <t>BL21</t> Gold, BL21 Codon (+) and BL21 <t>Rosetta</t> in the following growth media: Terrific Broth (TB, yellow), Auto-Induction medium (AI, blue) and LB medium (red). Lower panel, normalized ELIC band density from the western blot (Penta-histidine antibody) were plotted against E. coli strains grouped by growth media. 1c. The effect of the cell culture volume on ELIC expression levels and final culture biomass was studied for the three most promising E. coli strains: BL21 Gold, BL21 Codon (+) and BL21 Rosetta. 1d. The effect of Zn 2+ and Mg 2+ as channel negative modifiers or the competitive antagonist ACh on ELIC expression levels is represented as a bar plot.
    Induction Control E Bl21 De3, supplied by Millipore, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Millipore bl21 de3 t1r bacteria
    Optimization of conditions for the overexpression of ELIC 1a . A cartoon representation of ELIC highlighting the membrane spanning pore forming domain (red helices), the extracellular domain (yellow B-sheets), and the boundaries of the cell membrane in relation to the channel structure. 1b . Upper panel, a western blot analysis of the expression levels of the ELIC channel made in the E. coli strains: C41, C43, <t>BL21</t> Gold, BL21 Codon (+) and BL21 <t>Rosetta</t> in the following growth media: Terrific Broth (TB, yellow), Auto-Induction medium (AI, blue) and LB medium (red). Lower panel, normalized ELIC band density from the western blot (Penta-histidine antibody) were plotted against E. coli strains grouped by growth media. 1c. The effect of the cell culture volume on ELIC expression levels and final culture biomass was studied for the three most promising E. coli strains: BL21 Gold, BL21 Codon (+) and BL21 Rosetta. 1d. The effect of Zn 2+ and Mg 2+ as channel negative modifiers or the competitive antagonist ACh on ELIC expression levels is represented as a bar plot.
    Bl21 De3 T1r Bacteria, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore protein purification bl21
    Optimization of conditions for the overexpression of ELIC 1a . A cartoon representation of ELIC highlighting the membrane spanning pore forming domain (red helices), the extracellular domain (yellow B-sheets), and the boundaries of the cell membrane in relation to the channel structure. 1b . Upper panel, a western blot analysis of the expression levels of the ELIC channel made in the E. coli strains: C41, C43, <t>BL21</t> Gold, BL21 Codon (+) and BL21 <t>Rosetta</t> in the following growth media: Terrific Broth (TB, yellow), Auto-Induction medium (AI, blue) and LB medium (red). Lower panel, normalized ELIC band density from the western blot (Penta-histidine antibody) were plotted against E. coli strains grouped by growth media. 1c. The effect of the cell culture volume on ELIC expression levels and final culture biomass was studied for the three most promising E. coli strains: BL21 Gold, BL21 Codon (+) and BL21 Rosetta. 1d. The effect of Zn 2+ and Mg 2+ as channel negative modifiers or the competitive antagonist ACh on ELIC expression levels is represented as a bar plot.
    Protein Purification Bl21, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Millipore bl21 de3 plyss rosetta
    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
    Bl21 De3 Plyss Rosetta, supplied by Millipore, used in various techniques. Bioz Stars score: 82/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore bl21 de3 strain
    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
    Bl21 De3 Strain, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 237 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
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    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
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    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
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    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
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    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
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    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
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    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
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    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli <t>BL21</t> (DE3) <t>pLysS</t> <t>Rosetta</t> whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.
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    Image Search Results


    RP-HPLC separation of the heated cellular extract. Cellular extract of E. coli BL21 [DE3] (pMA24) were treated as described in Section 2.4 . Chromatogram was monitored at (A) λ = 220 nm and (B) λ = 280 nm. Peaks were collected and individually analyzed by MALDI-TOF MS.

    Journal: FEBS Open Bio

    Article Title: A new hybrid bacteriocin, Ent35-MccV, displays antimicrobial activity against pathogenic Gram-positive and Gram-negative bacteria

    doi: 10.1016/j.fob.2012.01.002

    Figure Lengend Snippet: RP-HPLC separation of the heated cellular extract. Cellular extract of E. coli BL21 [DE3] (pMA24) were treated as described in Section 2.4 . Chromatogram was monitored at (A) λ = 220 nm and (B) λ = 280 nm. Peaks were collected and individually analyzed by MALDI-TOF MS.

    Article Snippet: 2.3 Gene expression and bacteriocins purification for activity determinations Ent35–MccV was expressed from pMA24 in E. coli BL21 [DE3] (pLysS) grown in LB (Sigma) at 37 °C.

    Techniques: High Performance Liquid Chromatography, Mass Spectrometry

    Expression of UspF protein by an IPTG‐inducible E. coli BL21 (DE3) pLyS strain and purification of soluble fractions containing UspF protein. A – T0 total protein extract before induction; T2, total protein extract after induction (3 h); T3, total protein extract after induction (16–18 h). The arrow indicates position of the UspF protein (18.4 kDa). B – Purification of soluble fractions or UspF protein by affinity chromatography using a nickel‐containing resin. 1–Flow through; 2–30 m M , 3–50 m M , 4–100 m M ; 5–200 m M of imidazole.

    Journal: Protein Science : A Publication of the Protein Society

    Article Title: Characterization of the universal stress protein F from atypical enteropathogenic Escherichia coli and its prevalence in Enterobacteriaceae

    doi: 10.1002/pro.3038

    Figure Lengend Snippet: Expression of UspF protein by an IPTG‐inducible E. coli BL21 (DE3) pLyS strain and purification of soluble fractions containing UspF protein. A – T0 total protein extract before induction; T2, total protein extract after induction (3 h); T3, total protein extract after induction (16–18 h). The arrow indicates position of the UspF protein (18.4 kDa). B – Purification of soluble fractions or UspF protein by affinity chromatography using a nickel‐containing resin. 1–Flow through; 2–30 m M , 3–50 m M , 4–100 m M ; 5–200 m M of imidazole.

    Article Snippet: Bacterial strains and plasmids The following E. coli K12 strains were used: DH10b (Stratagene, USA) and BL21 (DE3) pLyS (Novagen, USA).

    Techniques: Expressing, Purification, Affinity Chromatography, Flow Cytometry

    A fly SUMO-ome: characterization and validation. A) Scheme for identifying Drosophila embryonic SUMO conjugates. SUMO conjugates were isolated by tandem affinity purification from transgenic fly embryos expressing (His) 6 -FLAG-SUMO. The initial purification step (Ni-NTA chromatography) was performed under denaturing conditions. To maximize the chance of detecting low abundance proteins in the complex protein mixture, the affinity-purified proteins were separated by SDS-PAGE, and the lane was cut into 20 evenly divided gel slices. Tryptic peptides extracted from each gel slice were analyzed by LC-MS/MS. B) A bacterial sumoylation assay. The Q SUMO vector, which encodes the mature form of SUMO (SUMO GG ) along with SAE1, SAE2, and Ubc9 expressed from separate T7/lac promoters, was used in combination with a vector expressing a GST-tagged candidate substrate. As a negative control, Q ΔGG , which expresses a conjugation defective form of SUMO (SUMO ΔGG ), was used in place of Q SUMO . C) Bacterial sumoylation assays were used to validate proteins identified in the proteomic screen as sumoylation substrates. GST-tagged candidate SUMO conjugates were expressed in BL21 cells co-transformed with Q SUMO or Q ΔGG vectors, purified using glutathione beads, and immunoblotted using antibodies against GST, SUMO, or poly-His (to detect 6xHis-tagged SUMO). GST by itself was not sumoylated in this assay. Black arrows point to the bands representing sumoylated proteins, and open arrow points to a non-specific reacting band. D) The eIF4E protein was purified from Drosophila S2 cells stably expressing FLAG-(His) 6 - tagged eIF4E using Ni-NTA beads under denaturing conditions. The resulting proteins were probed with anti-FLAG antibody in a Western blot. The cells were treated with SUMO or control YFP dsRNA for 3 days prior to cell lysis. In the control sample, the bands representing the sumoylated species (black arrows) have intensities that are 8.1% (top) and 12.9% (bottom) of the intensity of the band representing unmodified eIF4E (∼40 kDa), whereas in the SUMO knockdown sample, they are reduced to 1.8% (top) and 3.5% (bottom). Quantitation was performed using Quantity One 4.3.0 (BioRad).

    Journal: PLoS ONE

    Article Title: Genetic and Proteomic Evidence for Roles of Drosophila SUMO in Cell Cycle Control, Ras Signaling, and Early Pattern Formation

    doi: 10.1371/journal.pone.0005905

    Figure Lengend Snippet: A fly SUMO-ome: characterization and validation. A) Scheme for identifying Drosophila embryonic SUMO conjugates. SUMO conjugates were isolated by tandem affinity purification from transgenic fly embryos expressing (His) 6 -FLAG-SUMO. The initial purification step (Ni-NTA chromatography) was performed under denaturing conditions. To maximize the chance of detecting low abundance proteins in the complex protein mixture, the affinity-purified proteins were separated by SDS-PAGE, and the lane was cut into 20 evenly divided gel slices. Tryptic peptides extracted from each gel slice were analyzed by LC-MS/MS. B) A bacterial sumoylation assay. The Q SUMO vector, which encodes the mature form of SUMO (SUMO GG ) along with SAE1, SAE2, and Ubc9 expressed from separate T7/lac promoters, was used in combination with a vector expressing a GST-tagged candidate substrate. As a negative control, Q ΔGG , which expresses a conjugation defective form of SUMO (SUMO ΔGG ), was used in place of Q SUMO . C) Bacterial sumoylation assays were used to validate proteins identified in the proteomic screen as sumoylation substrates. GST-tagged candidate SUMO conjugates were expressed in BL21 cells co-transformed with Q SUMO or Q ΔGG vectors, purified using glutathione beads, and immunoblotted using antibodies against GST, SUMO, or poly-His (to detect 6xHis-tagged SUMO). GST by itself was not sumoylated in this assay. Black arrows point to the bands representing sumoylated proteins, and open arrow points to a non-specific reacting band. D) The eIF4E protein was purified from Drosophila S2 cells stably expressing FLAG-(His) 6 - tagged eIF4E using Ni-NTA beads under denaturing conditions. The resulting proteins were probed with anti-FLAG antibody in a Western blot. The cells were treated with SUMO or control YFP dsRNA for 3 days prior to cell lysis. In the control sample, the bands representing the sumoylated species (black arrows) have intensities that are 8.1% (top) and 12.9% (bottom) of the intensity of the band representing unmodified eIF4E (∼40 kDa), whereas in the SUMO knockdown sample, they are reduced to 1.8% (top) and 3.5% (bottom). Quantitation was performed using Quantity One 4.3.0 (BioRad).

    Article Snippet: The bacterial sumoylation assay A vector encoding a candidate SUMO conjugation target fused to GST was co-transformed into BL21 cells (Novagen) with either the QSUMO or the control QΔGG expression vectors.

    Techniques: Isolation, Affinity Purification, Transgenic Assay, Expressing, Purification, Chromatography, SDS Page, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Plasmid Preparation, Negative Control, Conjugation Assay, Transformation Assay, Stable Transfection, Western Blot, Lysis, Quantitation Assay

    Optimization of conditions for the overexpression of ELIC 1a . A cartoon representation of ELIC highlighting the membrane spanning pore forming domain (red helices), the extracellular domain (yellow B-sheets), and the boundaries of the cell membrane in relation to the channel structure. 1b . Upper panel, a western blot analysis of the expression levels of the ELIC channel made in the E. coli strains: C41, C43, BL21 Gold, BL21 Codon (+) and BL21 Rosetta in the following growth media: Terrific Broth (TB, yellow), Auto-Induction medium (AI, blue) and LB medium (red). Lower panel, normalized ELIC band density from the western blot (Penta-histidine antibody) were plotted against E. coli strains grouped by growth media. 1c. The effect of the cell culture volume on ELIC expression levels and final culture biomass was studied for the three most promising E. coli strains: BL21 Gold, BL21 Codon (+) and BL21 Rosetta. 1d. The effect of Zn 2+ and Mg 2+ as channel negative modifiers or the competitive antagonist ACh on ELIC expression levels is represented as a bar plot.

    Journal: Protein expression and purification

    Article Title: A cost-effective protocol for the over-expression and purification of fully-functional and more stable Erwinia chrisanthemi ligand-gated ion channel

    doi: 10.1016/j.pep.2017.03.006

    Figure Lengend Snippet: Optimization of conditions for the overexpression of ELIC 1a . A cartoon representation of ELIC highlighting the membrane spanning pore forming domain (red helices), the extracellular domain (yellow B-sheets), and the boundaries of the cell membrane in relation to the channel structure. 1b . Upper panel, a western blot analysis of the expression levels of the ELIC channel made in the E. coli strains: C41, C43, BL21 Gold, BL21 Codon (+) and BL21 Rosetta in the following growth media: Terrific Broth (TB, yellow), Auto-Induction medium (AI, blue) and LB medium (red). Lower panel, normalized ELIC band density from the western blot (Penta-histidine antibody) were plotted against E. coli strains grouped by growth media. 1c. The effect of the cell culture volume on ELIC expression levels and final culture biomass was studied for the three most promising E. coli strains: BL21 Gold, BL21 Codon (+) and BL21 Rosetta. 1d. The effect of Zn 2+ and Mg 2+ as channel negative modifiers or the competitive antagonist ACh on ELIC expression levels is represented as a bar plot.

    Article Snippet: E. coli strains: BL21 DE3 Gold, BL21 DE3 Codon (+), BL21 DE3 Rosetta (Novagen), C41, and C43 (Lucigen) were tested for their ability to express the MBP-ELIC fusion protein in sufficient quantity and quality.

    Techniques: Over Expression, Western Blot, Expressing, Cell Culture

    Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli BL21 (DE3) pLysS Rosetta whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.

    Journal: BMC Molecular Biology

    Article Title: Functional characterization of an alkaline exonuclease and single strand annealing protein from the SXT genetic element of Vibrio cholerae

    doi: 10.1186/1471-2199-12-16

    Figure Lengend Snippet: Purification of SXT-Exo and lambda-Exo, and determination of their multimericity by size exclusion chromatography . Panel A : Size exclusion chromatogram of purified SXT-Exo protein expressed from plasmid pEA1-1. Panel B : Size exclusion chromatogram of purified lambda-Exo protein expressed from plasmid pEE4. Panel C : 12% polyacrylamide gel (SDS-PAGE) analysis of the SXT-Exo purification procedure and purified SXT-Bet, SXT-Ssb, lambda-Bet and lambda-Exo proteins; lane 1: Benchmark protein ladder (Invitrogen); lane 2: pEA1-1/ E. coli BL21 (DE3) pLysS Rosetta whole cell extract immediately prior to induction; lane 3: whole cell extract 6 hours after induction with IPTG; lane 4: supernatant from cell extract 6 hours post induction; lane 5: purified SXT-Exo; lane 6: purified SXT-Bet expressed from pX28-1; lane 7: purified SXT-Ssb expressed from pSB2; lane 8: purified lambda-Bet expressed from p1DB; lane 9: purified lambda-Exo expressed from pEE4.

    Article Snippet: Protein expression was performed in E. coli BL21 (DE3) or BL21 (DE3) pLysS Rosetta (Novagen); inducing expression by the addition of isopropyl-1-thio-β-D-galactopyranoside (IPTG, USB) to between 0.1 to 0.5 mM; incubating cultures post-induction at 20-37°C.

    Techniques: Purification, Size-exclusion Chromatography, Plasmid Preparation, SDS Page

    Sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE) analysis of recombinant hG31P-P-113 expressed in E. coli BL21.

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli

    doi: 10.3390/molecules23040800

    Figure Lengend Snippet: Sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE) analysis of recombinant hG31P-P-113 expressed in E. coli BL21.

    Article Snippet: Materials and Methods The expression vector pET-28a and host BL21 (DE3) were purchased from Novagen (Merck Millipore, Burlington, MA, USA).

    Techniques: Polyacrylamide Gel Electrophoresis, Recombinant