bl21 de3 plyss  (Thermo Fisher)


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    Structured Review

    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: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 de3 plyss/product/Thermo Fisher
    Average 99 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    bl21 de3 plyss - by Bioz Stars, 2020-02
    99/100 stars

    Images

    1) Product Images from "PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS"

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

    Journal: Muscle & nerve

    doi: 10.1002/mus.23247

    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.
    Figure Legend 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.

    Techniques Used: Expressing, Purification

    Related Articles

    Clone Assay:

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: The amplified DNA fragment was cloned into pRSETA vector (Invitrogen, Inc.) for expressing wsx1 recombinant protein in a bacteria host. .. The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA).

    Article Title: Biological Activity of Recombinant Accessory Cholerae Enterotoxin (Ace) on Rabbit Ileal Loops and Antibacterial Assay
    Article Snippet: .. E. coli DH5α and BL21 (PlysS) were used for cloning and expression experiments (Invitrogen and Novagen, USA). .. Plasmid pET-28a+ (Novagen) was the expression vector.

    Article Title: Identification of Novel Adhesins of M. tuberculosis H37Rv Using Integrated Approach of Multiple Computational Algorithms and Experimental Analysis
    Article Snippet: Initially, cloning was done by PCR amplification of the entire open reading frame (ORF). .. Various E. coli strains, which could express toxic membrane proteins such as E. coli C43 (DE3), C41 (DE3) (Lucigen, USA), Rosetta (Novagen, Germany), BL21 (DE3) pLysS (Invitrogen, USA) were used for expression .

    Article Title: GW2 Functions as an E3 Ubiquitin Ligase for Rice Expansin-Like 1
    Article Snippet: Bacteria, Plant Materials, and Growth Conditions Escherichia coli strains DH5α, DH10B, and Top10 (Invitrogen, Waltham, MA, USA) were used for cloning and plasmid preparation. .. BL21/DE3 pLysS (Invitrogen) was used to express recombinant proteins.

    Centrifugation:

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG. .. The cell lysate was recovered by centrifugation at 4°C.

    Amplification:

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: The amplified DNA fragment was cloned into pRSETA vector (Invitrogen, Inc.) for expressing wsx1 recombinant protein in a bacteria host. .. The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA).

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: Purification of FtsZ protein The sequences coding N- and C-terminal S . aureus FtsZ were amplified by PCR, fused with EGFP and mCherry (red fluorescent protein), respectively, and inserted into pRSET vector (Invitrogen). .. The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG.

    Article Title: Identification of Novel Adhesins of M. tuberculosis H37Rv Using Integrated Approach of Multiple Computational Algorithms and Experimental Analysis
    Article Snippet: Initially, cloning was done by PCR amplification of the entire open reading frame (ORF). .. Various E. coli strains, which could express toxic membrane proteins such as E. coli C43 (DE3), C41 (DE3) (Lucigen, USA), Rosetta (Novagen, Germany), BL21 (DE3) pLysS (Invitrogen, USA) were used for expression .

    Filtration:

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG. .. After filtration, the lysate was purified using AKTAprime plus (GE Healthcare) installed in HisTrap HP and HiTrap Q HP columns (GE Healthcare) for Ni affinity and anion-exchange chromatography, respectively.

    Synthesized:

    Article Title: A biosynthetic pathway for a prominent class of microbiota-derived bile acids
    Article Snippet: E. coli TOP10 and BL21(DE3) pLysS chemically competent cells were purchased from Invitrogen. .. Oligonucleotide primers were synthesized by Elim Biopharm.

    Construct:

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: Mouse IL27 receptor wsx1 construct was purchased from Open Biosystems (Huntsville, AL). .. The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA).

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: A mutation was constructed at K175D to prevent further bundling of FtsZ in the N-terminal fragment. .. The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG.

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: Positive colonies on an agar-solidified LB medium were selected to isolate and confirm the recombinant construct by double enzyme digestion and sequencing, and then named pET28bMiANR1-1, 1-2, 1-3 respectively. .. To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively.

    Incubation:

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: .. The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG. .. After harvest, cells were lysed by sonication in a buffer consisting of 50 mM Tris-HCl (pH 8.0), 250 mM KCl and 20 mM imidazol supplemented with the following protease inhibitors: 50 μM bestatin (ACROS, Inc.), 800 μM AEBSF (Santa Cruz), 15 μM E-64, 20 μM pepstatinA, 7 μM phosphoramidon, and 10 μM leupeptin (Peptide Institute, INC) on ice.

    Article Title: Cross-Phosphorylation and Interaction between Src/FAK and MAPKAP5/PRAK in Early Focal Adhesions Controls Cell Motility
    Article Snippet: Expression of GST, GST-347PRAK, GST-PRAK-2, GST-paxillin, GST-HSP27 in BL21-pLysS (Invitrogen) was induced with 1 mM IPTG for 2–4 h at 30°C. .. Supernatants were incubated with GST-Bind resin (Novagen), and protein was eluted using GST Elution Buffer (50 mM Tris-Cl pH 8.0, 10 mM reduced glutathione).

    Expressing:

    Article Title: Suites of Terpene Synthases Explain Differential Terpenoid Production in Ginger and Turmeric Tissues
    Article Snippet: .. Expression of Terpene Synthases in E. coli and Yeast We used several E. coli strains for expression of terpene synthases; BL21 (DE3) pLysS (Invitrogen), BL21 CodonPlus (DE3) RIL (Stratagene), BL21 CodonPlus (DE3) RP (Stratagene), BL21 CodonPlus (DE3) RILP (Stratagene), Rosetta (Novagen), Rosetta2 (DE3) pLysS (Novagen), ArcticExpress (DE3) RIL (Stratagene), BL21-AI (Invitrogen), BL21-AI RIL, BL21 Star (DE3) (Invitrogen), BL21 Star (DE3) RIL and BL21 Star (DE3) pMevT pMBI RIL. .. The E. coli strain, BL21-AI RIL is BL21-AI plus the RIL plasmid from BL21 CodonPlus (DE3) RIL cells.

    Article Title: A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NF?B activity
    Article Snippet: Protein synthesis and purification TAT and TAT-RH plasmids were transformed into a BL21 (DE3) pLysS (Invitrogen) bacterial strain. .. 2 L of Luria Broth (LB) culture was grown overnight and then Isopropylthiogalactoside (IPTG, 100 μM, 3 hrs) was added to induce protein expression.

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: The amplified DNA fragment was cloned into pRSETA vector (Invitrogen, Inc.) for expressing wsx1 recombinant protein in a bacteria host. .. The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA).

    Article Title: Engineering a Virus-Like Particle as an Antigenic Platform for a Pfs47-Targeted Malaria Transmission-Blocking Vaccine
    Article Snippet: Paragraph title: Expression of AP205-SpyCatcher ... Briefly, BL21(DE3) pLysS chemically competent E. coli cells (ThermoFisher) or E. coli OverExpress™ C41(DE3) (Lucigen) were transformed with pEt17b-AP205-SpyCatcher or pET24-AP205-SpyCatcher and inoculated onto LB agar plates with 100 µg/mL ampicillin and 34 µg/mL chloramphenicol.

    Article Title: Biological Activity of Recombinant Accessory Cholerae Enterotoxin (Ace) on Rabbit Ileal Loops and Antibacterial Assay
    Article Snippet: .. E. coli DH5α and BL21 (PlysS) were used for cloning and expression experiments (Invitrogen and Novagen, USA). .. Plasmid pET-28a+ (Novagen) was the expression vector.

    Article Title: Cross-Phosphorylation and Interaction between Src/FAK and MAPKAP5/PRAK in Early Focal Adhesions Controls Cell Motility
    Article Snippet: .. Expression of GST, GST-347PRAK, GST-PRAK-2, GST-paxillin, GST-HSP27 in BL21-pLysS (Invitrogen) was induced with 1 mM IPTG for 2–4 h at 30°C. .. Bacteria were collected and lysed in GST binding buffer (50 mM Tris-Cl pH 7.4, 1 mM EDTA, 100 mM NaCl) by freeze/thaw followed by sonication.

    Article Title: Identification of Novel Adhesins of M. tuberculosis H37Rv Using Integrated Approach of Multiple Computational Algorithms and Experimental Analysis
    Article Snippet: .. Various E. coli strains, which could express toxic membrane proteins such as E. coli C43 (DE3), C41 (DE3) (Lucigen, USA), Rosetta (Novagen, Germany), BL21 (DE3) pLysS (Invitrogen, USA) were used for expression . .. Finally, we chose to remove the initial hydrophobic regions in the selected genes, which either encoded for signal peptide or for transmembrane helix and cloned the rest of the ORF and expressed the protein and purified for further analysis.

    Article Title: Expression and characterization of highly antigenic domains of chicken anemia virus viral VP2 and VP3 subunit proteins in a recombinant E. coli for sero-diagnostic applications
    Article Snippet: Bacterial strains and inoculation Two E. coli strains, BL21 (DE3) and BL21(DE3)-pLysS were purchased from Invitrogen Life Technologies (Carlsbad, CA) and Stratagene (La Jolla, CA), respectively. .. The overnight culture were then inoculated into 50 ml LB medium and grown at 37°C for 3 h by which time the optical density of the culture had reach 0.5 of OD600 and could be used for competent cell preparation and protein expression.

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: .. To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively. ..

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    Article Snippet: Paragraph title: Expression, Purification, and Renaturation of Recombinant scFv-DAF ... 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.

    Bradford Assay:

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    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. .. The protein elution was analyzed by 12% sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE), and protein concentration was calculated using the Bradford assay.

    Transformation Assay:

    Article Title: A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NF?B activity
    Article Snippet: .. Protein synthesis and purification TAT and TAT-RH plasmids were transformed into a BL21 (DE3) pLysS (Invitrogen) bacterial strain. .. 2 L of Luria Broth (LB) culture was grown overnight and then Isopropylthiogalactoside (IPTG, 100 μM, 3 hrs) was added to induce protein expression.

    Article Title: Engineering a Virus-Like Particle as an Antigenic Platform for a Pfs47-Targeted Malaria Transmission-Blocking Vaccine
    Article Snippet: .. Briefly, BL21(DE3) pLysS chemically competent E. coli cells (ThermoFisher) or E. coli OverExpress™ C41(DE3) (Lucigen) were transformed with pEt17b-AP205-SpyCatcher or pET24-AP205-SpyCatcher and inoculated onto LB agar plates with 100 µg/mL ampicillin and 34 µg/mL chloramphenicol. .. Ten transformed BL21 colonies were picked and cultured in 10 separate tubes containing 4 mL LB supplemented with 100 µg/mL ampicillin and 34 µg/mL chloramphenicol O/N at 37 °C.

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    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. .. Cells were induced with 1 mM isopropyl- β - d -thiogalactoside (IPTG) for 4 h according to the plasmid manufacturer’s instructions (Novagen).

    Chromatography:

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG. .. After filtration, the lysate was purified using AKTAprime plus (GE Healthcare) installed in HisTrap HP and HiTrap Q HP columns (GE Healthcare) for Ni affinity and anion-exchange chromatography, respectively.

    Cell Culture:

    Article Title: Engineering a Virus-Like Particle as an Antigenic Platform for a Pfs47-Targeted Malaria Transmission-Blocking Vaccine
    Article Snippet: Briefly, BL21(DE3) pLysS chemically competent E. coli cells (ThermoFisher) or E. coli OverExpress™ C41(DE3) (Lucigen) were transformed with pEt17b-AP205-SpyCatcher or pET24-AP205-SpyCatcher and inoculated onto LB agar plates with 100 µg/mL ampicillin and 34 µg/mL chloramphenicol. .. Ten transformed BL21 colonies were picked and cultured in 10 separate tubes containing 4 mL LB supplemented with 100 µg/mL ampicillin and 34 µg/mL chloramphenicol O/N at 37 °C.

    Article Title: Biological Activity of Recombinant Accessory Cholerae Enterotoxin (Ace) on Rabbit Ileal Loops and Antibacterial Assay
    Article Snippet: E. coli DH5α and BL21 (PlysS) were used for cloning and expression experiments (Invitrogen and Novagen, USA). .. Bacteria were cultured in LB broth or on agar (Merck, Germany) with or without 30 µg kanamycin/ml (Sigma, USA).

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively. .. Then IPTG was added to a final concentration of 0.5 mm and further cultured at 30 °C and 220× g rpm overnight.

    SDS Page:

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA). .. The column purified wsx1 protein was further purified with SDS-PAGE to remove any contaminated proteins.

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    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. .. The protein elution was analyzed by 12% sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE), and protein concentration was calculated using the Bradford assay.

    Protein Concentration:

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    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. .. The protein elution was analyzed by 12% sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE), and protein concentration was calculated using the Bradford assay.

    Polymerase Chain Reaction:

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: Wsx1 encoding DNA was amplified from the purchased wsx1 clone via polymerase chain reaction (PCR) using forward and reverse primers Svm198F: 5′-CGGATCCATGAACCGGCTCCGGGTT-3′ and SVM199R: 5′-TCAGACTAGAAGGCCCAGCTC-3′, respectively. .. The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA).

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: Purification of FtsZ protein The sequences coding N- and C-terminal S . aureus FtsZ were amplified by PCR, fused with EGFP and mCherry (red fluorescent protein), respectively, and inserted into pRSET vector (Invitrogen). .. The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG.

    Article Title: Identification of Novel Adhesins of M. tuberculosis H37Rv Using Integrated Approach of Multiple Computational Algorithms and Experimental Analysis
    Article Snippet: Initially, cloning was done by PCR amplification of the entire open reading frame (ORF). .. Various E. coli strains, which could express toxic membrane proteins such as E. coli C43 (DE3), C41 (DE3) (Lucigen, USA), Rosetta (Novagen, Germany), BL21 (DE3) pLysS (Invitrogen, USA) were used for expression .

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: Then PCR was carried out to amplify respective ORFs and the products were digested with restriction enzyme NcoI and XhoI, followed by purification with the DNA fragment Purification Kit (QIA quick ® PCR pufication kit). .. To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively.

    Sonication:

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG. .. After harvest, cells were lysed by sonication in a buffer consisting of 50 mM Tris-HCl (pH 8.0), 250 mM KCl and 20 mM imidazol supplemented with the following protease inhibitors: 50 μM bestatin (ACROS, Inc.), 800 μM AEBSF (Santa Cruz), 15 μM E-64, 20 μM pepstatinA, 7 μM phosphoramidon, and 10 μM leupeptin (Peptide Institute, INC) on ice.

    Article Title: Cross-Phosphorylation and Interaction between Src/FAK and MAPKAP5/PRAK in Early Focal Adhesions Controls Cell Motility
    Article Snippet: Expression of GST, GST-347PRAK, GST-PRAK-2, GST-paxillin, GST-HSP27 in BL21-pLysS (Invitrogen) was induced with 1 mM IPTG for 2–4 h at 30°C. .. Bacteria were collected and lysed in GST binding buffer (50 mM Tris-Cl pH 7.4, 1 mM EDTA, 100 mM NaCl) by freeze/thaw followed by sonication.

    Injection:

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA). .. Prior to immunizing the rabbits (2.5 kg per rabbit) with the homogenized protein-gel slurry, 100 μg homogenized protein-gel slurry was mixed with 1 mL Complete Freund adjuvant by syringe thoroughly and injected subcutaneously.

    Recombinant:

    Article Title: A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NF?B activity
    Article Snippet: Protein synthesis and purification TAT and TAT-RH plasmids were transformed into a BL21 (DE3) pLysS (Invitrogen) bacterial strain. .. For protein purification we used denaturing conditions to recover all the recombinant proteins from bacterial inclusion bodies (Lysis buffer: 8 M Urea, 100 mM NaCl, 20 mM Hepes pH 8).

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: The amplified DNA fragment was cloned into pRSETA vector (Invitrogen, Inc.) for expressing wsx1 recombinant protein in a bacteria host. .. The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA).

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: .. To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively. ..

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    Article Snippet: Paragraph title: Expression, Purification, and Renaturation of Recombinant scFv-DAF ... 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.

    Article Title: GW2 Functions as an E3 Ubiquitin Ligase for Rice Expansin-Like 1
    Article Snippet: .. BL21/DE3 pLysS (Invitrogen) was used to express recombinant proteins. .. Wild-type, mutant, and transgenic rice were grown at 26 °C under long-day conditions (16 h light/8 h dark) in a greenhouse or field.

    Molecular Weight:

    Article Title: Cross-Phosphorylation and Interaction between Src/FAK and MAPKAP5/PRAK in Early Focal Adhesions Controls Cell Motility
    Article Snippet: Expression of GST, GST-347PRAK, GST-PRAK-2, GST-paxillin, GST-HSP27 in BL21-pLysS (Invitrogen) was induced with 1 mM IPTG for 2–4 h at 30°C. .. Eluted protein was dialyzed 1:1000 into PBS at 4°C using Slide-a-Lyzer cartridges (Pierce) or de-salted and concentrated using Amicon ultra centrifugal filters with 10 kDa molecular weight cutoff (Millipore).

    Nucleic Acid Electrophoresis:

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    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. .. The protein elution was analyzed by 12% sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE), and protein concentration was calculated using the Bradford assay.

    Mutagenesis:

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: A mutation was constructed at K175D to prevent further bundling of FtsZ in the N-terminal fragment. .. The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG.

    Article Title: GW2 Functions as an E3 Ubiquitin Ligase for Rice Expansin-Like 1
    Article Snippet: BL21/DE3 pLysS (Invitrogen) was used to express recombinant proteins. .. Wild-type, mutant, and transgenic rice were grown at 26 °C under long-day conditions (16 h light/8 h dark) in a greenhouse or field.

    Purification:

    Article Title: A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NF?B activity
    Article Snippet: .. Protein synthesis and purification TAT and TAT-RH plasmids were transformed into a BL21 (DE3) pLysS (Invitrogen) bacterial strain. .. 2 L of Luria Broth (LB) culture was grown overnight and then Isopropylthiogalactoside (IPTG, 100 μM, 3 hrs) was added to induce protein expression.

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: .. The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA). .. The column purified wsx1 protein was further purified with SDS-PAGE to remove any contaminated proteins.

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: Paragraph title: Purification of FtsZ protein ... The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG.

    Article Title: Identification of Novel Adhesins of M. tuberculosis H37Rv Using Integrated Approach of Multiple Computational Algorithms and Experimental Analysis
    Article Snippet: Paragraph title: Protein Expression and purification ... Various E. coli strains, which could express toxic membrane proteins such as E. coli C43 (DE3), C41 (DE3) (Lucigen, USA), Rosetta (Novagen, Germany), BL21 (DE3) pLysS (Invitrogen, USA) were used for expression .

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: The purified cDNA were then ligated into the pET28b (Novagen, San Diego, CA, USA) vector that had already been digested by NcoI and XhoI. .. To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively.

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    Article Snippet: Paragraph title: Expression, Purification, and Renaturation of Recombinant scFv-DAF ... 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.

    Protein Purification:

    Article Title: A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NF?B activity
    Article Snippet: Protein synthesis and purification TAT and TAT-RH plasmids were transformed into a BL21 (DE3) pLysS (Invitrogen) bacterial strain. .. For protein purification we used denaturing conditions to recover all the recombinant proteins from bacterial inclusion bodies (Lysis buffer: 8 M Urea, 100 mM NaCl, 20 mM Hepes pH 8).

    Article Title: Cross-Phosphorylation and Interaction between Src/FAK and MAPKAP5/PRAK in Early Focal Adhesions Controls Cell Motility
    Article Snippet: Paragraph title: Protein purification ... Expression of GST, GST-347PRAK, GST-PRAK-2, GST-paxillin, GST-HSP27 in BL21-pLysS (Invitrogen) was induced with 1 mM IPTG for 2–4 h at 30°C.

    Sequencing:

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: Positive colonies on an agar-solidified LB medium were selected to isolate and confirm the recombinant construct by double enzyme digestion and sequencing, and then named pET28bMiANR1-1, 1-2, 1-3 respectively. .. To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively.

    Positron Emission Tomography:

    Article Title: Biological Activity of Recombinant Accessory Cholerae Enterotoxin (Ace) on Rabbit Ileal Loops and Antibacterial Assay
    Article Snippet: E. coli DH5α and BL21 (PlysS) were used for cloning and expression experiments (Invitrogen and Novagen, USA). .. Plasmid pET-28a+ (Novagen) was the expression vector.

    Affinity Purification:

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: This ligation led to the fusion of cDNA to the C-terminal containing a His-tag-coding sequence in the vector, which is necessary for Ni-NIA affinity purification. .. To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively.

    Plasmid Preparation:

    Article Title: Suites of Terpene Synthases Explain Differential Terpenoid Production in Ginger and Turmeric Tissues
    Article Snippet: Expression of Terpene Synthases in E. coli and Yeast We used several E. coli strains for expression of terpene synthases; BL21 (DE3) pLysS (Invitrogen), BL21 CodonPlus (DE3) RIL (Stratagene), BL21 CodonPlus (DE3) RP (Stratagene), BL21 CodonPlus (DE3) RILP (Stratagene), Rosetta (Novagen), Rosetta2 (DE3) pLysS (Novagen), ArcticExpress (DE3) RIL (Stratagene), BL21-AI (Invitrogen), BL21-AI RIL, BL21 Star (DE3) (Invitrogen), BL21 Star (DE3) RIL and BL21 Star (DE3) pMevT pMBI RIL. .. The E. coli strain, BL21-AI RIL is BL21-AI plus the RIL plasmid from BL21 CodonPlus (DE3) RIL cells.

    Article Title: A biosynthetic pathway for a prominent class of microbiota-derived bile acids
    Article Snippet: E. coli TOP10 and BL21(DE3) pLysS chemically competent cells were purchased from Invitrogen. .. Plasmid pET28b was purchased from Novagen.

    Article Title: IL12 and IL27 sequential gene therapy via intramuscular electroporation delivery for eliminating distal aggressive tumors 1
    Article Snippet: The amplified DNA fragment was cloned into pRSETA vector (Invitrogen, Inc.) for expressing wsx1 recombinant protein in a bacteria host. .. The bacteria host used in this study was BL21 DE3 plysS and the wsx1 protein was purified using ProBond Purification System under denaturing conditions (Invitrogen, Carlsbad, CA).

    Article Title: Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis
    Article Snippet: Purification of FtsZ protein The sequences coding N- and C-terminal S . aureus FtsZ were amplified by PCR, fused with EGFP and mCherry (red fluorescent protein), respectively, and inserted into pRSET vector (Invitrogen). .. The proteins of FtsZK175D _N-terminal-EGFP and FtsZ_C-terminal-mCherry were expressed in E . coli , BL21 pLysS (Invitrogen), and incubated at 15°C for 20 hours with 0.5 mM IPTG.

    Article Title: Biological Activity of Recombinant Accessory Cholerae Enterotoxin (Ace) on Rabbit Ileal Loops and Antibacterial Assay
    Article Snippet: E. coli DH5α and BL21 (PlysS) were used for cloning and expression experiments (Invitrogen and Novagen, USA). .. Plasmid pET-28a+ (Novagen) was the expression vector.

    Article Title: Identification of Novel Adhesins of M. tuberculosis H37Rv Using Integrated Approach of Multiple Computational Algorithms and Experimental Analysis
    Article Snippet: The vector pET28a was from Novagen, USA. .. Various E. coli strains, which could express toxic membrane proteins such as E. coli C43 (DE3), C41 (DE3) (Lucigen, USA), Rosetta (Novagen, Germany), BL21 (DE3) pLysS (Invitrogen, USA) were used for expression .

    Article Title: Serodiagnostic comparison of enzyme-linked immunosorbent assay and surface plasmon resonance for the detection of antibody to Porcine circovirus type 2
    Article Snippet: Escherichia coli strains JM109 and BL21(DE3)pLysS were purchased from Invitrogen (Carlsbad, California, USA). .. The pRSET vector (Invitrogen), which has been described previously , was used to produce the 6X histidine-tagged protein.

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: .. To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively. ..

    Article Title: PROTECTIVE EFFECT OF ScFv-DAF FUSION PROTEIN ON THE COMPLEMENT ATTACK TO ACETYLCHOLINE RECEPTOR: A POSSIBLE OPTION FOR TREATMENT OF MYASTHENIA GRAVIS
    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. .. Cells were induced with 1 mM isopropyl- β - d -thiogalactoside (IPTG) for 4 h according to the plasmid manufacturer’s instructions (Novagen).

    Article Title: GW2 Functions as an E3 Ubiquitin Ligase for Rice Expansin-Like 1
    Article Snippet: Bacteria, Plant Materials, and Growth Conditions Escherichia coli strains DH5α, DH10B, and Top10 (Invitrogen, Waltham, MA, USA) were used for cloning and plasmid preparation. .. BL21/DE3 pLysS (Invitrogen) was used to express recombinant proteins.

    Binding Assay:

    Article Title: Cross-Phosphorylation and Interaction between Src/FAK and MAPKAP5/PRAK in Early Focal Adhesions Controls Cell Motility
    Article Snippet: Expression of GST, GST-347PRAK, GST-PRAK-2, GST-paxillin, GST-HSP27 in BL21-pLysS (Invitrogen) was induced with 1 mM IPTG for 2–4 h at 30°C. .. Bacteria were collected and lysed in GST binding buffer (50 mM Tris-Cl pH 7.4, 1 mM EDTA, 100 mM NaCl) by freeze/thaw followed by sonication.

    Transgenic Assay:

    Article Title: GW2 Functions as an E3 Ubiquitin Ligase for Rice Expansin-Like 1
    Article Snippet: BL21/DE3 pLysS (Invitrogen) was used to express recombinant proteins. .. Wild-type, mutant, and transgenic rice were grown at 26 °C under long-day conditions (16 h light/8 h dark) in a greenhouse or field.

    Concentration Assay:

    Article Title: Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L.
    Article Snippet: To study protein expression, the recombinant vectors pET28bMiANR1-1, 1-2, 1-3, pET28bMiANR1-1, 1-2, 1-3 and an empty vector pET28b(+) (control) were introduced into ultra-competent cells of the Bl21 (De3) plysS (Invitrogen, Carlsbad, CA, USA) strain, respectively. .. Then IPTG was added to a final concentration of 0.5 mm and further cultured at 30 °C and 220× g rpm overnight.

    Lysis:

    Article Title: A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NF?B activity
    Article Snippet: Protein synthesis and purification TAT and TAT-RH plasmids were transformed into a BL21 (DE3) pLysS (Invitrogen) bacterial strain. .. For protein purification we used denaturing conditions to recover all the recombinant proteins from bacterial inclusion bodies (Lysis buffer: 8 M Urea, 100 mM NaCl, 20 mM Hepes pH 8).

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