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Merck KGaA e coli bl21
SDS-PAGE (a) and western blot (b) analyses of the recombinant BirA protein expressed via pET24-BirA vector in <t>BL21</t> E. coli cells by addition of IPTG to the final concentration of 1 mM. (a) Lane 1: Protein molecular weight marker (Unstained protein molecular weight marker, Fermentas, Lithuania). Different induced colonies (lane 3–7) showed the same size of BirA (36.6 kDa) expression compared with un-induced sample (lane 2) (b) Western blot analysis of recombinant BirA protein by anti-His antibody specific for C-terminal polyhistidine tag indicated the specific 36.6 kDa protein band. Lane 1: Protein marker (PrestainedProtein Ladder, CinnaGen, Iran) Lane 2: Un-induced bacterial lysate. Lane 3: Recombinant BirA protein (36.6 kDa). Arrows indicate the target protein bands
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1) Product Images from "Isolation, cloning, and expression of E. coli BirA gene for biotinylation applications"

Article Title: Isolation, cloning, and expression of E. coli BirA gene for biotinylation applications

Journal: Advanced Biomedical Research

doi: 10.4103/2277-9175.161576

SDS-PAGE (a) and western blot (b) analyses of the recombinant BirA protein expressed via pET24-BirA vector in BL21 E. coli cells by addition of IPTG to the final concentration of 1 mM. (a) Lane 1: Protein molecular weight marker (Unstained protein molecular weight marker, Fermentas, Lithuania). Different induced colonies (lane 3–7) showed the same size of BirA (36.6 kDa) expression compared with un-induced sample (lane 2) (b) Western blot analysis of recombinant BirA protein by anti-His antibody specific for C-terminal polyhistidine tag indicated the specific 36.6 kDa protein band. Lane 1: Protein marker (PrestainedProtein Ladder, CinnaGen, Iran) Lane 2: Un-induced bacterial lysate. Lane 3: Recombinant BirA protein (36.6 kDa). Arrows indicate the target protein bands
Figure Legend Snippet: SDS-PAGE (a) and western blot (b) analyses of the recombinant BirA protein expressed via pET24-BirA vector in BL21 E. coli cells by addition of IPTG to the final concentration of 1 mM. (a) Lane 1: Protein molecular weight marker (Unstained protein molecular weight marker, Fermentas, Lithuania). Different induced colonies (lane 3–7) showed the same size of BirA (36.6 kDa) expression compared with un-induced sample (lane 2) (b) Western blot analysis of recombinant BirA protein by anti-His antibody specific for C-terminal polyhistidine tag indicated the specific 36.6 kDa protein band. Lane 1: Protein marker (PrestainedProtein Ladder, CinnaGen, Iran) Lane 2: Un-induced bacterial lysate. Lane 3: Recombinant BirA protein (36.6 kDa). Arrows indicate the target protein bands

Techniques Used: SDS Page, Western Blot, Recombinant, Plasmid Preparation, Concentration Assay, Molecular Weight, Marker, Expressing

2) Product Images from "Cloning, expression and purification of the factor H binding protein and its interaction with factor H"

Article Title: Cloning, expression and purification of the factor H binding protein and its interaction with factor H

Journal: Iranian Journal of Microbiology

doi:

SDS-PAGE and western blot Analysis of the purified recombinant protein after purification: (A) Lane 1, purified protein; 2; Lane M, molecular weight marker; Lane 3, bacterial lysate after 3h incubation. (B) Lane 1, purified protein; Lane M, molecular weight marker; Lane 2, BL21 cell lysate.
Figure Legend Snippet: SDS-PAGE and western blot Analysis of the purified recombinant protein after purification: (A) Lane 1, purified protein; 2; Lane M, molecular weight marker; Lane 3, bacterial lysate after 3h incubation. (B) Lane 1, purified protein; Lane M, molecular weight marker; Lane 2, BL21 cell lysate.

Techniques Used: SDS Page, Western Blot, Purification, Recombinant, Molecular Weight, Marker, Incubation

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Clone Assay:

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Centrifugation:

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Amplification:

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Filtration:

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TA Cloning:

Article Title: Isolation, cloning, and expression of E. coli BirA gene for biotinylation applications
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Incubation:

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Expressing:

Article Title: Comparative Transcription Profiling and In-Depth Characterization of Plasmid-Based and Plasmid-Free Escherichia coli Expression Systems under Production Conditions
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Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
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Article Title: The Amino Acid Exchange R28E in Ciliary Neurotrophic Factor (CNTF) Abrogates Interleukin-6 Receptor-dependent but Retains CNTF Receptor-dependent Signaling via Glycoprotein 130 (gp130)/Leukemia Inhibitory Factor Receptor (LIFR) *
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Article Title: Cloning, expression and purification of the factor H binding protein and its interaction with factor H
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Article Title: Influence of N-glycosylation on effector functions and thermal stability of glycoengineered IgG1 monoclonal antibody with homogeneous glycoforms
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Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
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Transformation Assay:

Article Title: Comparative Transcription Profiling and In-Depth Characterization of Plasmid-Based and Plasmid-Free Escherichia coli Expression Systems under Production Conditions
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Article Title: Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs
Article Snippet: .. The positive recombinants were confirmed by PCR and DNA sequencing and then transformed into E. coli BL21(DE3) (Merck Millipore) for expression. .. Overnight cultures of E. coli BL21(DE3) strains carrying the recombinant plasmids were used to inoculate 1 to 6 liters fresh 2YT broth, grown to an OD595 of 0.6 at 37°C in broth supplemented with 100 μg/ml kanamycin, and then induced with 1 mM IPTG (isopropyl-β- d -1-thiogalactopyranoside; Sigma) at 37°C for 2, 4, and 24 h. Protein expression was checked by SDS-PAGE using whole-cell lysates.

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59
Article Snippet: .. PCR product comprising ILY coding gene lacking the putative signal sequence (aa residues 1–33) was cloned into pET28a(+) vector (Novagen/Merck KGaA, Darmstadt, Germany) and transformed into E. coli BL21(DE3) strain (Merck KGaA, Darmstadt, Germany). .. Recombinant N -terminally-hexahistidine-tagged ILY was purified from 2 L of culture using affinity chromatography on 6BCL-IDA Ni-sepharose (GE Healthcare, Helsinki, Finland) and ion-exchange chromatography on SP sepharose FF (GE Healthcare, Helsinki, Finland) according to manufacturer’s recommendations.

Article Title: A distal phenylalanine clamp in a hydrophobic channel controls the substrate specificity in the quorum-quenching metallo-\u03b3-lactonase (AiiA) from Bacillus thuringiensis
Article Snippet: .. Briefly, E. coli BL21(DE3) cells (Merck, Damstadt, Germany) transformed with the pMAL-t-AiiA vector were grown and induced with 0.5 mM IPTG. .. AiiA was purified as described with an additional gel filtration step ( , ).

Article Title: Influence of N-glycosylation on effector functions and thermal stability of glycoengineered IgG1 monoclonal antibody with homogeneous glycoforms
Article Snippet: .. Recombinant expression of GST-EndoS2 and GST-EndoS2-D184M Each plasmid was transformed into E. coli BL21(DE3) (Merck Millipore, Singles™ Competent Cells, Cat # 70235–3) and grown on an agar LB (Nacalai Tesque, Cat # 20066–95) plate containing 20 μg/mL kanamycin (Wako, Cat # 115–00342) overnight at 37 °C. .. Each recombinant clone was selected and cultured in a liquid LB medium containing 20 μg/mL kanamycin for six hours at 30 °C to obtain research cell banks (RCB), maintained as stocks in glycerol (Nacalai Tesque, Cat # 17018–83).

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: .. For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL). .. The precipitate was dissolved in 5 ml lysis buffer (8 M urea, 100 mM NaH2PO4, 10 mMTris-HCl, pH: 8.0) and following centrifugation at 12000 rpm for 30 minutes.

Derivative Assay:

Article Title: Comparative Transcription Profiling and In-Depth Characterization of Plasmid-Based and Plasmid-Free Escherichia coli Expression Systems under Production Conditions
Article Snippet: Experiments were performed using E. coli BL21(DE3) (Merck KGaA, Darmstadt, Germany). .. BL21(DE3) was transformed with pET30aSOD, yielding BL21(DE3)(pET30aSOD). pET30aSOD was derived by subcloning of the Homo sapiens superoxide dismutase 1 gene (GenBank accession no. ) from pET11aSOD, described elsewhere (SOD, superoxide dismutase) ( ).

Chromatography:

Article Title: Biophysical Properties of Intrinsically Disordered p130Cas Substrate Domain -- Implication in Mechanosensing
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Ligation:

Article Title: Direct induction of microtubule branching by microtubule nucleation factor SSNA1
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Protease Inhibitor:

Article Title: Biophysical Properties of Intrinsically Disordered p130Cas Substrate Domain -- Implication in Mechanosensing
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Cell Culture:

Article Title: Isolation, cloning, and expression of E. coli BirA gene for biotinylation applications
Article Snippet: E. coli BL21 (DE3) and pET-24a(+) plasmid (Merck KGaA, Darmstadt, Germany) were used as expression host and vector, respectively. .. Bacterial strains were cultured in Luria-Bertani medium (supplemented with 50 μg/mL kanamycin when required).

Article Title: Discovery of Sanggenon G as a natural cell-permeable small-molecular weight inhibitor of X-linked inhibitor of apoptosis protein (XIAP)
Article Snippet: Protein expression was performed in Escherichia coli BL21(DE3)pLysS (Merck Millipore, Germany). .. E. coli carrying the pET30a(+)-XIAP-BIR3 plasmid were cultured overnight in 25 ml of LB-medium with 50 μg/ml kanamycin at 37 °C.

Article Title: Influence of N-glycosylation on effector functions and thermal stability of glycoengineered IgG1 monoclonal antibody with homogeneous glycoforms
Article Snippet: Recombinant expression of GST-EndoS2 and GST-EndoS2-D184M Each plasmid was transformed into E. coli BL21(DE3) (Merck Millipore, Singles™ Competent Cells, Cat # 70235–3) and grown on an agar LB (Nacalai Tesque, Cat # 20066–95) plate containing 20 μg/mL kanamycin (Wako, Cat # 115–00342) overnight at 37 °C. .. Each recombinant clone was selected and cultured in a liquid LB medium containing 20 μg/mL kanamycin for six hours at 30 °C to obtain research cell banks (RCB), maintained as stocks in glycerol (Nacalai Tesque, Cat # 17018–83).

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: Briefly, a single colony of transformed E. coli BL21 (DE3) containing pET-15b/lenA was cultured on 5 ml Luria Bertani (LB) broth containing ampicillin (100 μg/mL) and incubated at 37°C overnight (about 16 h). .. For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL).

DNA Sequencing:

Article Title: Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs
Article Snippet: .. The positive recombinants were confirmed by PCR and DNA sequencing and then transformed into E. coli BL21(DE3) (Merck Millipore) for expression. .. Overnight cultures of E. coli BL21(DE3) strains carrying the recombinant plasmids were used to inoculate 1 to 6 liters fresh 2YT broth, grown to an OD595 of 0.6 at 37°C in broth supplemented with 100 μg/ml kanamycin, and then induced with 1 mM IPTG (isopropyl-β- d -1-thiogalactopyranoside; Sigma) at 37°C for 2, 4, and 24 h. Protein expression was checked by SDS-PAGE using whole-cell lysates.

Sequencing:

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59
Article Snippet: .. PCR product comprising ILY coding gene lacking the putative signal sequence (aa residues 1–33) was cloned into pET28a(+) vector (Novagen/Merck KGaA, Darmstadt, Germany) and transformed into E. coli BL21(DE3) strain (Merck KGaA, Darmstadt, Germany). .. Recombinant N -terminally-hexahistidine-tagged ILY was purified from 2 L of culture using affinity chromatography on 6BCL-IDA Ni-sepharose (GE Healthcare, Helsinki, Finland) and ion-exchange chromatography on SP sepharose FF (GE Healthcare, Helsinki, Finland) according to manufacturer’s recommendations.

Sonication:

Article Title: Direct induction of microtubule branching by microtubule nucleation factor SSNA1
Article Snippet: The proteins were expressed in E. coli BL21(DE3) (Merck, Darmstadt, Germany) by inducing with 0.4 mM IPTG (Carl Roth, Karlsruhe, Germany) for overnight at 18 °C. .. Cells were sonicated in lysis buffer (50 mM Na-phosphate buffer pH 7.5, 150 mM NaCl, 10% (v/v) glycerol, 5mM β-mercaptoethanol) supplemented with protease inhibitors (1 mM Pepstatin A, 1mM AEBSF and 1mM Leupeptin) and clarified.

Article Title: Biophysical Properties of Intrinsically Disordered p130Cas Substrate Domain -- Implication in Mechanosensing
Article Snippet: Cloning, expression, and purification of recombinant CasSD Mouse CasSD was produced as a tobacco etch virus (TEV) protease-cleavable C-terminal His6 -tagged protein in the E. coli BL21(DE3) Rosetta2 strain (Merck Biosciences, Darmstadt, Germany). .. Cell suspension in a lysis buffer (50 mM potassium phosphate pH 7.8, 300 mM potassium chloride, protease inhibitor cocktail VII (Merck Biosciences, Darmstadt, Germany) was sonicated and centrifuged to obtain a cleared cell lysate.

Binding Assay:

Article Title: A distal phenylalanine clamp in a hydrophobic channel controls the substrate specificity in the quorum-quenching metallo-\u03b3-lactonase (AiiA) from Bacillus thuringiensis
Article Snippet: Bacillus thuringiensis AHL lactonase (AiiA) was expressed and purified as a TEV (Tobacco Etch Virus) protease cleavable fusion to maltose binding protein according to the procedure reported previously unless noted otherwise ( , ). .. Briefly, E. coli BL21(DE3) cells (Merck, Damstadt, Germany) transformed with the pMAL-t-AiiA vector were grown and induced with 0.5 mM IPTG.

Nucleic Acid Electrophoresis:

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: The pellet and supernatant were transferred to separate tubes, and the cell pellets were suspended in 100 μl sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) sample loading buffer and were subjected to 12.5% SDS-PAGE. .. For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL).

Ion Exchange Chromatography:

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59
Article Snippet: PCR product comprising ILY coding gene lacking the putative signal sequence (aa residues 1–33) was cloned into pET28a(+) vector (Novagen/Merck KGaA, Darmstadt, Germany) and transformed into E. coli BL21(DE3) strain (Merck KGaA, Darmstadt, Germany). .. Recombinant N -terminally-hexahistidine-tagged ILY was purified from 2 L of culture using affinity chromatography on 6BCL-IDA Ni-sepharose (GE Healthcare, Helsinki, Finland) and ion-exchange chromatography on SP sepharose FF (GE Healthcare, Helsinki, Finland) according to manufacturer’s recommendations.

Mutagenesis:

Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
Article Snippet: .. 2.2 Production and purification of the mutant and wild-type Sfβgly Expression vectors pET46 coding for the wild-type and mutant Sfβgly were separately introduced in E. coli BL21(DE3) (Merck Millipore; Darmstad, Germany) by heat shock following manufacturer's procedures. ..

Isolation:

Article Title: Isolation, cloning, and expression of E. coli BirA gene for biotinylation applications
Article Snippet: Bacterial strains and plasmids E. coli DH5α, a derivative of E. coli K-12,[ ] was used both as target strain for isolation of the BirA DNA fragment and also as the primary host for plasmid manipulations and cloning steps. pTG19-T cloning vector (Vivantis, Selangor DE, Malaysia) was used for TA cloning steps. .. E. coli BL21 (DE3) and pET-24a(+) plasmid (Merck KGaA, Darmstadt, Germany) were used as expression host and vector, respectively.

Subcloning:

Article Title: Comparative Transcription Profiling and In-Depth Characterization of Plasmid-Based and Plasmid-Free Escherichia coli Expression Systems under Production Conditions
Article Snippet: Experiments were performed using E. coli BL21(DE3) (Merck KGaA, Darmstadt, Germany). .. BL21(DE3) was transformed with pET30aSOD, yielding BL21(DE3)(pET30aSOD). pET30aSOD was derived by subcloning of the Homo sapiens superoxide dismutase 1 gene (GenBank accession no. ) from pET11aSOD, described elsewhere (SOD, superoxide dismutase) ( ).

Purification:

Article Title: UBE2B is implicated in myofibrillar protein loss in catabolic C2C12 myotubes) UBE2B is implicated in myofibrillar protein loss in catabolic C2C12 myotubes
Article Snippet: GST pulldown assays GST‐MuRF1 and UBE2B were expressed in Escherichia coli BL21(DE3) (Merck, Darmstadt, Germany) using 100 μM IPTG. .. GST‐MuRF1 were then purified with Glutathione‐Sepharose 4B beads (GE Healthcare) according to the manufacturers' instructions and used as a 50% slurry for subsequent GST‐pulldown experiments.

Article Title: Discovery of Sanggenon G as a natural cell-permeable small-molecular weight inhibitor of X-linked inhibitor of apoptosis protein (XIAP)
Article Snippet: Paragraph title: Production and purification of recombinant BIR3 protein ... Protein expression was performed in Escherichia coli BL21(DE3)pLysS (Merck Millipore, Germany).

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59
Article Snippet: PCR product comprising ILY coding gene lacking the putative signal sequence (aa residues 1–33) was cloned into pET28a(+) vector (Novagen/Merck KGaA, Darmstadt, Germany) and transformed into E. coli BL21(DE3) strain (Merck KGaA, Darmstadt, Germany). .. Recombinant N -terminally-hexahistidine-tagged ILY was purified from 2 L of culture using affinity chromatography on 6BCL-IDA Ni-sepharose (GE Healthcare, Helsinki, Finland) and ion-exchange chromatography on SP sepharose FF (GE Healthcare, Helsinki, Finland) according to manufacturer’s recommendations.

Article Title: A distal phenylalanine clamp in a hydrophobic channel controls the substrate specificity in the quorum-quenching metallo-\u03b3-lactonase (AiiA) from Bacillus thuringiensis
Article Snippet: Paragraph title: Protein expression and purification ... Briefly, E. coli BL21(DE3) cells (Merck, Damstadt, Germany) transformed with the pMAL-t-AiiA vector were grown and induced with 0.5 mM IPTG.

Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
Article Snippet: .. 2.2 Production and purification of the mutant and wild-type Sfβgly Expression vectors pET46 coding for the wild-type and mutant Sfβgly were separately introduced in E. coli BL21(DE3) (Merck Millipore; Darmstad, Germany) by heat shock following manufacturer's procedures. ..

Article Title: The Amino Acid Exchange R28E in Ciliary Neurotrophic Factor (CNTF) Abrogates Interleukin-6 Receptor-dependent but Retains CNTF Receptor-dependent Signaling via Glycoprotein 130 (gp130)/Leukemia Inhibitory Factor Receptor (LIFR) *
Article Snippet: Paragraph title: Expression, Purification, and Renaturation of CV-1 to CV-5, CLC, IC-7, IL-6, and CNTF ... Expression of CV-1 to CV-5, CLC, IC-7, IL-6, and CNTF was performed in Escherichia coli BL21(DE3) (Merck KGaA).

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: .. For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL). .. The precipitate was dissolved in 5 ml lysis buffer (8 M urea, 100 mM NaH2PO4, 10 mMTris-HCl, pH: 8.0) and following centrifugation at 12000 rpm for 30 minutes.

Article Title: Direct induction of microtubule branching by microtubule nucleation factor SSNA1
Article Snippet: Paragraph title: Protein preparation and purification ... The proteins were expressed in E. coli BL21(DE3) (Merck, Darmstadt, Germany) by inducing with 0.4 mM IPTG (Carl Roth, Karlsruhe, Germany) for overnight at 18 °C.

Article Title: Biophysical Properties of Intrinsically Disordered p130Cas Substrate Domain -- Implication in Mechanosensing
Article Snippet: .. Cloning, expression, and purification of recombinant CasSD Mouse CasSD was produced as a tobacco etch virus (TEV) protease-cleavable C-terminal His6 -tagged protein in the E. coli BL21(DE3) Rosetta2 strain (Merck Biosciences, Darmstadt, Germany). .. Induction of the gene expression was achieved by 37°C incubation for three hours after addition of 400 µM isopropyl-β-d -thiogalactopyranoside (IPTG) to LB culture.

Polymerase Chain Reaction:

Article Title: Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs
Article Snippet: .. The positive recombinants were confirmed by PCR and DNA sequencing and then transformed into E. coli BL21(DE3) (Merck Millipore) for expression. .. Overnight cultures of E. coli BL21(DE3) strains carrying the recombinant plasmids were used to inoculate 1 to 6 liters fresh 2YT broth, grown to an OD595 of 0.6 at 37°C in broth supplemented with 100 μg/ml kanamycin, and then induced with 1 mM IPTG (isopropyl-β- d -1-thiogalactopyranoside; Sigma) at 37°C for 2, 4, and 24 h. Protein expression was checked by SDS-PAGE using whole-cell lysates.

Article Title: Discovery of Sanggenon G as a natural cell-permeable small-molecular weight inhibitor of X-linked inhibitor of apoptosis protein (XIAP)
Article Snippet: 2.8 Production and purification of recombinant BIR3 protein DNA encoding BIR3 of human XIAP protein was amplified by PCR and cloned into the pET30a(+) vector (Merck Millipore, Germany) via the NdeI and SalI restriction sites. .. Protein expression was performed in Escherichia coli BL21(DE3)pLysS (Merck Millipore, Germany).

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59
Article Snippet: .. PCR product comprising ILY coding gene lacking the putative signal sequence (aa residues 1–33) was cloned into pET28a(+) vector (Novagen/Merck KGaA, Darmstadt, Germany) and transformed into E. coli BL21(DE3) strain (Merck KGaA, Darmstadt, Germany). .. Recombinant N -terminally-hexahistidine-tagged ILY was purified from 2 L of culture using affinity chromatography on 6BCL-IDA Ni-sepharose (GE Healthcare, Helsinki, Finland) and ion-exchange chromatography on SP sepharose FF (GE Healthcare, Helsinki, Finland) according to manufacturer’s recommendations.

Affinity Chromatography:

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59
Article Snippet: PCR product comprising ILY coding gene lacking the putative signal sequence (aa residues 1–33) was cloned into pET28a(+) vector (Novagen/Merck KGaA, Darmstadt, Germany) and transformed into E. coli BL21(DE3) strain (Merck KGaA, Darmstadt, Germany). .. Recombinant N -terminally-hexahistidine-tagged ILY was purified from 2 L of culture using affinity chromatography on 6BCL-IDA Ni-sepharose (GE Healthcare, Helsinki, Finland) and ion-exchange chromatography on SP sepharose FF (GE Healthcare, Helsinki, Finland) according to manufacturer’s recommendations.

Article Title: Direct induction of microtubule branching by microtubule nucleation factor SSNA1
Article Snippet: The proteins were expressed in E. coli BL21(DE3) (Merck, Darmstadt, Germany) by inducing with 0.4 mM IPTG (Carl Roth, Karlsruhe, Germany) for overnight at 18 °C. .. Soluble fraction was purified by Ni-NTA affinity chromatography.

Recombinant:

Article Title: Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs
Article Snippet: The positive recombinants were confirmed by PCR and DNA sequencing and then transformed into E. coli BL21(DE3) (Merck Millipore) for expression. .. Overnight cultures of E. coli BL21(DE3) strains carrying the recombinant plasmids were used to inoculate 1 to 6 liters fresh 2YT broth, grown to an OD595 of 0.6 at 37°C in broth supplemented with 100 μg/ml kanamycin, and then induced with 1 mM IPTG (isopropyl-β- d -1-thiogalactopyranoside; Sigma) at 37°C for 2, 4, and 24 h. Protein expression was checked by SDS-PAGE using whole-cell lysates.

Article Title: Discovery of Sanggenon G as a natural cell-permeable small-molecular weight inhibitor of X-linked inhibitor of apoptosis protein (XIAP)
Article Snippet: Paragraph title: Production and purification of recombinant BIR3 protein ... Protein expression was performed in Escherichia coli BL21(DE3)pLysS (Merck Millipore, Germany).

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59
Article Snippet: PCR product comprising ILY coding gene lacking the putative signal sequence (aa residues 1–33) was cloned into pET28a(+) vector (Novagen/Merck KGaA, Darmstadt, Germany) and transformed into E. coli BL21(DE3) strain (Merck KGaA, Darmstadt, Germany). .. Recombinant N -terminally-hexahistidine-tagged ILY was purified from 2 L of culture using affinity chromatography on 6BCL-IDA Ni-sepharose (GE Healthcare, Helsinki, Finland) and ion-exchange chromatography on SP sepharose FF (GE Healthcare, Helsinki, Finland) according to manufacturer’s recommendations.

Article Title: Cloning, expression and purification of the factor H binding protein and its interaction with factor H
Article Snippet: Paragraph title: Expression of the recombinant fhbp gene. ... E. coli BL21 (DE3) was used for protein expression as host with 50μg/μl kanamycin (Merck,Germany) in LB medium for selection, 0.5 mM IPTG (Isopropyl-beta-D-thiogalacto-pyranoside) (Merck,Germany) as inducer.

Article Title: Influence of N-glycosylation on effector functions and thermal stability of glycoengineered IgG1 monoclonal antibody with homogeneous glycoforms
Article Snippet: .. Recombinant expression of GST-EndoS2 and GST-EndoS2-D184M Each plasmid was transformed into E. coli BL21(DE3) (Merck Millipore, Singles™ Competent Cells, Cat # 70235–3) and grown on an agar LB (Nacalai Tesque, Cat # 20066–95) plate containing 20 μg/mL kanamycin (Wako, Cat # 115–00342) overnight at 37 °C. .. Each recombinant clone was selected and cultured in a liquid LB medium containing 20 μg/mL kanamycin for six hours at 30 °C to obtain research cell banks (RCB), maintained as stocks in glycerol (Nacalai Tesque, Cat # 17018–83).

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: .. For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL). .. The precipitate was dissolved in 5 ml lysis buffer (8 M urea, 100 mM NaH2PO4, 10 mMTris-HCl, pH: 8.0) and following centrifugation at 12000 rpm for 30 minutes.

Article Title: Biophysical Properties of Intrinsically Disordered p130Cas Substrate Domain -- Implication in Mechanosensing
Article Snippet: .. Cloning, expression, and purification of recombinant CasSD Mouse CasSD was produced as a tobacco etch virus (TEV) protease-cleavable C-terminal His6 -tagged protein in the E. coli BL21(DE3) Rosetta2 strain (Merck Biosciences, Darmstadt, Germany). .. Induction of the gene expression was achieved by 37°C incubation for three hours after addition of 400 µM isopropyl-β-d -thiogalactopyranoside (IPTG) to LB culture.

SDS Page:

Article Title: Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs
Article Snippet: The positive recombinants were confirmed by PCR and DNA sequencing and then transformed into E. coli BL21(DE3) (Merck Millipore) for expression. .. Overnight cultures of E. coli BL21(DE3) strains carrying the recombinant plasmids were used to inoculate 1 to 6 liters fresh 2YT broth, grown to an OD595 of 0.6 at 37°C in broth supplemented with 100 μg/ml kanamycin, and then induced with 1 mM IPTG (isopropyl-β- d -1-thiogalactopyranoside; Sigma) at 37°C for 2, 4, and 24 h. Protein expression was checked by SDS-PAGE using whole-cell lysates.

Article Title: A distal phenylalanine clamp in a hydrophobic channel controls the substrate specificity in the quorum-quenching metallo-\u03b3-lactonase (AiiA) from Bacillus thuringiensis
Article Snippet: Briefly, E. coli BL21(DE3) cells (Merck, Damstadt, Germany) transformed with the pMAL-t-AiiA vector were grown and induced with 0.5 mM IPTG. .. Fractions containing monomeric AiiA, identified by the elution volume and SDS-PAGE, were pooled and concentrated.

Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
Article Snippet: 2.2 Production and purification of the mutant and wild-type Sfβgly Expression vectors pET46 coding for the wild-type and mutant Sfβgly were separately introduced in E. coli BL21(DE3) (Merck Millipore; Darmstad, Germany) by heat shock following manufacturer's procedures. .. The purity of the Sfβgly samples were checked by using SDS-PAGE .

Article Title: Cloning, expression and purification of the factor H binding protein and its interaction with factor H
Article Snippet: E. coli BL21 (DE3) was used for protein expression as host with 50μg/μl kanamycin (Merck,Germany) in LB medium for selection, 0.5 mM IPTG (Isopropyl-beta-D-thiogalacto-pyranoside) (Merck,Germany) as inducer. .. The cells were harvested, treated with lysis buffer (50 mM Tris base, 10% glycerol, 0.1%Triton X-100) (Merck, Germany) and the lysate was analyzed by SDS-PAGE and the quantity of the expressed protein was estimated by comparing the intensity of the protein bands.

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: The pellet and supernatant were transferred to separate tubes, and the cell pellets were suspended in 100 μl sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) sample loading buffer and were subjected to 12.5% SDS-PAGE. .. For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL).

Plasmid Preparation:

Article Title: Comparative Transcription Profiling and In-Depth Characterization of Plasmid-Based and Plasmid-Free Escherichia coli Expression Systems under Production Conditions
Article Snippet: Experiments were performed using E. coli BL21(DE3) (Merck KGaA, Darmstadt, Germany). .. Linear double-stranded DNA (dsDNA) cartridges, containing the expression unit from pET30a < SOD > fused to a chloramphenicol acetyltransferase resistance gene ( cat ), were integrated into the bacterial chromosome at the att Tn 7 site of E. coli BL21(DE3), which carries the pSIM6 helper plasmid, as described by Sharan et al. ( ).

Article Title: Isolation, cloning, and expression of E. coli BirA gene for biotinylation applications
Article Snippet: .. E. coli BL21 (DE3) and pET-24a(+) plasmid (Merck KGaA, Darmstadt, Germany) were used as expression host and vector, respectively. .. Bacterial strains were cultured in Luria-Bertani medium (supplemented with 50 μg/mL kanamycin when required).

Article Title: Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs
Article Snippet: The PCR products of candidate genes were cloned into the pET-30 Ek/LIC vector (Merck Millipore), and fusion plasmids were transformed into E. coli NovaBlue (Merck Millipore) according to the manufacturer's instructions. .. The positive recombinants were confirmed by PCR and DNA sequencing and then transformed into E. coli BL21(DE3) (Merck Millipore) for expression.

Article Title: Discovery of Sanggenon G as a natural cell-permeable small-molecular weight inhibitor of X-linked inhibitor of apoptosis protein (XIAP)
Article Snippet: 2.8 Production and purification of recombinant BIR3 protein DNA encoding BIR3 of human XIAP protein was amplified by PCR and cloned into the pET30a(+) vector (Merck Millipore, Germany) via the NdeI and SalI restriction sites. .. Protein expression was performed in Escherichia coli BL21(DE3)pLysS (Merck Millipore, Germany).

Article Title: The Cytolytic Activity of Vaginolysin Strictly Depends on Cholesterol and Is Potentiated by Human CD59
Article Snippet: .. PCR product comprising ILY coding gene lacking the putative signal sequence (aa residues 1–33) was cloned into pET28a(+) vector (Novagen/Merck KGaA, Darmstadt, Germany) and transformed into E. coli BL21(DE3) strain (Merck KGaA, Darmstadt, Germany). .. Recombinant N -terminally-hexahistidine-tagged ILY was purified from 2 L of culture using affinity chromatography on 6BCL-IDA Ni-sepharose (GE Healthcare, Helsinki, Finland) and ion-exchange chromatography on SP sepharose FF (GE Healthcare, Helsinki, Finland) according to manufacturer’s recommendations.

Article Title: A distal phenylalanine clamp in a hydrophobic channel controls the substrate specificity in the quorum-quenching metallo-\u03b3-lactonase (AiiA) from Bacillus thuringiensis
Article Snippet: .. Briefly, E. coli BL21(DE3) cells (Merck, Damstadt, Germany) transformed with the pMAL-t-AiiA vector were grown and induced with 0.5 mM IPTG. .. AiiA was purified as described with an additional gel filtration step ( , ).

Article Title: Influence of N-glycosylation on effector functions and thermal stability of glycoengineered IgG1 monoclonal antibody with homogeneous glycoforms
Article Snippet: .. Recombinant expression of GST-EndoS2 and GST-EndoS2-D184M Each plasmid was transformed into E. coli BL21(DE3) (Merck Millipore, Singles™ Competent Cells, Cat # 70235–3) and grown on an agar LB (Nacalai Tesque, Cat # 20066–95) plate containing 20 μg/mL kanamycin (Wako, Cat # 115–00342) overnight at 37 °C. .. Each recombinant clone was selected and cultured in a liquid LB medium containing 20 μg/mL kanamycin for six hours at 30 °C to obtain research cell banks (RCB), maintained as stocks in glycerol (Nacalai Tesque, Cat # 17018–83).

Article Title: Selection of High Producers From Combinatorial Libraries for the Production of Recombinant Proteins in Escherichia coli and Vibrio natriegens
Article Snippet: Strains and Growth Conditions Escherichia coli NEB 10-beta (NEB, Ipswich, Massachusetts, USA) was used for cloning and plasmid amplification. .. E. coli BL21(DE3) (Merck, Darmstadt, Germany) and V. natriegens Vmax Express (SGI-DNA, La Jolla, California, USA) served as expression hosts.

Positron Emission Tomography:

Article Title: Isolation, cloning, and expression of E. coli BirA gene for biotinylation applications
Article Snippet: .. E. coli BL21 (DE3) and pET-24a(+) plasmid (Merck KGaA, Darmstadt, Germany) were used as expression host and vector, respectively. .. Bacterial strains were cultured in Luria-Bertani medium (supplemented with 50 μg/mL kanamycin when required).

Article Title: Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs
Article Snippet: The PCR products of candidate genes were cloned into the pET-30 Ek/LIC vector (Merck Millipore), and fusion plasmids were transformed into E. coli NovaBlue (Merck Millipore) according to the manufacturer's instructions. .. The positive recombinants were confirmed by PCR and DNA sequencing and then transformed into E. coli BL21(DE3) (Merck Millipore) for expression.

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: Briefly, a single colony of transformed E. coli BL21 (DE3) containing pET-15b/lenA was cultured on 5 ml Luria Bertani (LB) broth containing ampicillin (100 μg/mL) and incubated at 37°C overnight (about 16 h). .. For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL).

Selection:

Article Title: Cloning, expression and purification of the factor H binding protein and its interaction with factor H
Article Snippet: .. E. coli BL21 (DE3) was used for protein expression as host with 50μg/μl kanamycin (Merck,Germany) in LB medium for selection, 0.5 mM IPTG (Isopropyl-beta-D-thiogalacto-pyranoside) (Merck,Germany) as inducer. .. The cells were harvested, treated with lysis buffer (50 mM Tris base, 10% glycerol, 0.1%Triton X-100) (Merck, Germany) and the lysate was analyzed by SDS-PAGE and the quantity of the expressed protein was estimated by comparing the intensity of the protein bands.

Article Title: Selection of High Producers From Combinatorial Libraries for the Production of Recombinant Proteins in Escherichia coli and Vibrio natriegens
Article Snippet: E. coli BL21(DE3) (Merck, Darmstadt, Germany) and V. natriegens Vmax Express (SGI-DNA, La Jolla, California, USA) served as expression hosts. .. The following antibiotics were used for plasmid selection: ampicillin (100 μg/mL), kanamycin (35 μg/mL) and spectinomycin (E. coli 65 μg/mL, V. natriegens 50 μg/mL).

Produced:

Article Title: Biophysical Properties of Intrinsically Disordered p130Cas Substrate Domain -- Implication in Mechanosensing
Article Snippet: .. Cloning, expression, and purification of recombinant CasSD Mouse CasSD was produced as a tobacco etch virus (TEV) protease-cleavable C-terminal His6 -tagged protein in the E. coli BL21(DE3) Rosetta2 strain (Merck Biosciences, Darmstadt, Germany). .. Induction of the gene expression was achieved by 37°C incubation for three hours after addition of 400 µM isopropyl-β-d -thiogalactopyranoside (IPTG) to LB culture.

Concentration Assay:

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: Then a final concentration of 1 mM of IPTG (isopropyl-β-D-thiogalactopyranoside) [Sigma-Aldrich] was added to bacterial cultures and the cells were grown for 4 hours at 37°C with constant shaking. .. For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL).

Lysis:

Article Title: Cloning, expression and purification of the factor H binding protein and its interaction with factor H
Article Snippet: E. coli BL21 (DE3) was used for protein expression as host with 50μg/μl kanamycin (Merck,Germany) in LB medium for selection, 0.5 mM IPTG (Isopropyl-beta-D-thiogalacto-pyranoside) (Merck,Germany) as inducer. .. The cells were harvested, treated with lysis buffer (50 mM Tris base, 10% glycerol, 0.1%Triton X-100) (Merck, Germany) and the lysate was analyzed by SDS-PAGE and the quantity of the expressed protein was estimated by comparing the intensity of the protein bands.

Article Title: Vaccine potential of LenA and LcpA proteins of Leptospira interrogans in combination with Escherichia coli heat-labile enterotoxin, B subunit (LTB)
Article Snippet: For purification of the recombinant protein, transformed E. coli BL21 (DE3) overexpressed in 500 ml Luria Bertani broth (Merck, Germany) containing ampicillin (100 μg/mL). .. The precipitate was dissolved in 5 ml lysis buffer (8 M urea, 100 mM NaH2PO4, 10 mMTris-HCl, pH: 8.0) and following centrifugation at 12000 rpm for 30 minutes.

Article Title: Direct induction of microtubule branching by microtubule nucleation factor SSNA1
Article Snippet: The proteins were expressed in E. coli BL21(DE3) (Merck, Darmstadt, Germany) by inducing with 0.4 mM IPTG (Carl Roth, Karlsruhe, Germany) for overnight at 18 °C. .. Cells were sonicated in lysis buffer (50 mM Na-phosphate buffer pH 7.5, 150 mM NaCl, 10% (v/v) glycerol, 5mM β-mercaptoethanol) supplemented with protease inhibitors (1 mM Pepstatin A, 1mM AEBSF and 1mM Leupeptin) and clarified.

Article Title: Biophysical Properties of Intrinsically Disordered p130Cas Substrate Domain -- Implication in Mechanosensing
Article Snippet: Cloning, expression, and purification of recombinant CasSD Mouse CasSD was produced as a tobacco etch virus (TEV) protease-cleavable C-terminal His6 -tagged protein in the E. coli BL21(DE3) Rosetta2 strain (Merck Biosciences, Darmstadt, Germany). .. Cell suspension in a lysis buffer (50 mM potassium phosphate pH 7.8, 300 mM potassium chloride, protease inhibitor cocktail VII (Merck Biosciences, Darmstadt, Germany) was sonicated and centrifuged to obtain a cleared cell lysate.

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  • 93
    Merck KGaA e coli bl21
    SDS-PAGE (a) and western blot (b) analyses of the recombinant BirA protein expressed via pET24-BirA vector in <t>BL21</t> E. coli cells by addition of IPTG to the final concentration of 1 mM. (a) Lane 1: Protein molecular weight marker (Unstained protein molecular weight marker, Fermentas, Lithuania). Different induced colonies (lane 3–7) showed the same size of BirA (36.6 kDa) expression compared with un-induced sample (lane 2) (b) Western blot analysis of recombinant BirA protein by anti-His antibody specific for C-terminal polyhistidine tag indicated the specific 36.6 kDa protein band. Lane 1: Protein marker (PrestainedProtein Ladder, CinnaGen, Iran) Lane 2: Un-induced bacterial lysate. Lane 3: Recombinant BirA protein (36.6 kDa). Arrows indicate the target protein bands
    E Coli Bl21, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21/product/Merck KGaA
    Average 93 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 - by Bioz Stars, 2020-01
    93/100 stars
      Buy from Supplier

    80
    Merck KGaA bl21 de3 plyss strain
    Small Scale expression analysis from pBDDP-SPR3 constructs. GTPases were expressed in the <t>Bl21</t> (DE3) <t>pLysS</t> strain of E. coli in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate. A 1 ml sample of cells was collected before induction and at the time of cell harvesting. This allowed the assessment of total cell protein by SDS-PAGE, and the visualisation of over-expressed target protein. A 40 ml sample of expression culture was applied to a small scale CoNTA IMAC purification to demonstrate the presence of soluble his-tagged protein. (As reported by others, CoNTA spin column purification has proved particularly useful in this respect and gives a consistently clear indication of levels and integrity of recombinant proteins [20] .) In all three cases there was significant soluble expression of the GTPase allowing progression to FPLC scale purification (H 8 -H 8 -KRas 1–169, 24.8 kDa; H 8 -H 8 -Rac1-2-177, 26.2 kDa; H 8 -H 8 -RalB, 25.5 kDa).
    Bl21 De3 Plyss Strain, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 80/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 de3 plyss strain/product/Merck KGaA
    Average 80 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    bl21 de3 plyss strain - by Bioz Stars, 2020-01
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    83
    Merck KGaA ms2 coat protein sodium dodecyl sulfate polyacrylamide gel electrophoresis sds page
    Sodium dodecyl sulfate-polyacrylamide gel electrophoresis <t>(SDS-PAGE</t> – left part of the picture) and Western blot ( right part of the picture) results. 1,6, E. coli BL21 (DE3) negative control; 2,5, IPTG-non-induced culture; 3,4, IPTG-induced culture. 13 kDa <t>MS2</t> coat protein was massively produced in the IPTG-induced culture. M, marker Spectra multicolor broad range protein ladder (Fermentas), the marker values are in kDa.
    Ms2 Coat Protein Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis Sds Page, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 83/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ms2 coat protein sodium dodecyl sulfate polyacrylamide gel electrophoresis sds page/product/Merck KGaA
    Average 83 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ms2 coat protein sodium dodecyl sulfate polyacrylamide gel electrophoresis sds page - by Bioz Stars, 2020-01
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    SDS-PAGE (a) and western blot (b) analyses of the recombinant BirA protein expressed via pET24-BirA vector in BL21 E. coli cells by addition of IPTG to the final concentration of 1 mM. (a) Lane 1: Protein molecular weight marker (Unstained protein molecular weight marker, Fermentas, Lithuania). Different induced colonies (lane 3–7) showed the same size of BirA (36.6 kDa) expression compared with un-induced sample (lane 2) (b) Western blot analysis of recombinant BirA protein by anti-His antibody specific for C-terminal polyhistidine tag indicated the specific 36.6 kDa protein band. Lane 1: Protein marker (PrestainedProtein Ladder, CinnaGen, Iran) Lane 2: Un-induced bacterial lysate. Lane 3: Recombinant BirA protein (36.6 kDa). Arrows indicate the target protein bands

    Journal: Advanced Biomedical Research

    Article Title: Isolation, cloning, and expression of E. coli BirA gene for biotinylation applications

    doi: 10.4103/2277-9175.161576

    Figure Lengend Snippet: SDS-PAGE (a) and western blot (b) analyses of the recombinant BirA protein expressed via pET24-BirA vector in BL21 E. coli cells by addition of IPTG to the final concentration of 1 mM. (a) Lane 1: Protein molecular weight marker (Unstained protein molecular weight marker, Fermentas, Lithuania). Different induced colonies (lane 3–7) showed the same size of BirA (36.6 kDa) expression compared with un-induced sample (lane 2) (b) Western blot analysis of recombinant BirA protein by anti-His antibody specific for C-terminal polyhistidine tag indicated the specific 36.6 kDa protein band. Lane 1: Protein marker (PrestainedProtein Ladder, CinnaGen, Iran) Lane 2: Un-induced bacterial lysate. Lane 3: Recombinant BirA protein (36.6 kDa). Arrows indicate the target protein bands

    Article Snippet: E. coli BL21 (DE3) and pET-24a(+) plasmid (Merck KGaA, Darmstadt, Germany) were used as expression host and vector, respectively.

    Techniques: SDS Page, Western Blot, Recombinant, Plasmid Preparation, Concentration Assay, Molecular Weight, Marker, Expressing

    SDS-PAGE and western blot Analysis of the purified recombinant protein after purification: (A) Lane 1, purified protein; 2; Lane M, molecular weight marker; Lane 3, bacterial lysate after 3h incubation. (B) Lane 1, purified protein; Lane M, molecular weight marker; Lane 2, BL21 cell lysate.

    Journal: Iranian Journal of Microbiology

    Article Title: Cloning, expression and purification of the factor H binding protein and its interaction with factor H

    doi:

    Figure Lengend Snippet: SDS-PAGE and western blot Analysis of the purified recombinant protein after purification: (A) Lane 1, purified protein; 2; Lane M, molecular weight marker; Lane 3, bacterial lysate after 3h incubation. (B) Lane 1, purified protein; Lane M, molecular weight marker; Lane 2, BL21 cell lysate.

    Article Snippet: E. coli BL21 (DE3) was used for protein expression as host with 50μg/μl kanamycin (Merck,Germany) in LB medium for selection, 0.5 mM IPTG (Isopropyl-beta-D-thiogalacto-pyranoside) (Merck,Germany) as inducer.

    Techniques: SDS Page, Western Blot, Purification, Recombinant, Molecular Weight, Marker, Incubation

    Small Scale expression analysis from pBDDP-SPR3 constructs. GTPases were expressed in the Bl21 (DE3) pLysS strain of E. coli in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate. A 1 ml sample of cells was collected before induction and at the time of cell harvesting. This allowed the assessment of total cell protein by SDS-PAGE, and the visualisation of over-expressed target protein. A 40 ml sample of expression culture was applied to a small scale CoNTA IMAC purification to demonstrate the presence of soluble his-tagged protein. (As reported by others, CoNTA spin column purification has proved particularly useful in this respect and gives a consistently clear indication of levels and integrity of recombinant proteins [20] .) In all three cases there was significant soluble expression of the GTPase allowing progression to FPLC scale purification (H 8 -H 8 -KRas 1–169, 24.8 kDa; H 8 -H 8 -Rac1-2-177, 26.2 kDa; H 8 -H 8 -RalB, 25.5 kDa).

    Journal: Protein Expression and Purification

    Article Title: A fully automated procedure for the parallel, multidimensional purification and nucleotide loading of the human GTPases KRas, Rac1 and RalB

    doi: 10.1016/j.pep.2017.01.010

    Figure Lengend Snippet: Small Scale expression analysis from pBDDP-SPR3 constructs. GTPases were expressed in the Bl21 (DE3) pLysS strain of E. coli in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate. A 1 ml sample of cells was collected before induction and at the time of cell harvesting. This allowed the assessment of total cell protein by SDS-PAGE, and the visualisation of over-expressed target protein. A 40 ml sample of expression culture was applied to a small scale CoNTA IMAC purification to demonstrate the presence of soluble his-tagged protein. (As reported by others, CoNTA spin column purification has proved particularly useful in this respect and gives a consistently clear indication of levels and integrity of recombinant proteins [20] .) In all three cases there was significant soluble expression of the GTPase allowing progression to FPLC scale purification (H 8 -H 8 -KRas 1–169, 24.8 kDa; H 8 -H 8 -Rac1-2-177, 26.2 kDa; H 8 -H 8 -RalB, 25.5 kDa).

    Article Snippet: 2.2 Expression of the CDC25 GEF domain of SOS1 and G-domains of KRas 4B, Rac1 and RalB in pBDDP-SPR3 Plasmids were transformed into the BL21 (DE3) pLysS strain of E. coli (Merck Millipore) and the transformed cells were cultured overnight at 37 °C in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate (Melford Laboratories Ltd).

    Techniques: Expressing, Construct, Cell Harvesting, SDS Page, Purification, Recombinant, Fast Protein Liquid Chromatography

    Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE – left part of the picture) and Western blot ( right part of the picture) results. 1,6, E. coli BL21 (DE3) negative control; 2,5, IPTG-non-induced culture; 3,4, IPTG-induced culture. 13 kDa MS2 coat protein was massively produced in the IPTG-induced culture. M, marker Spectra multicolor broad range protein ladder (Fermentas), the marker values are in kDa.

    Journal: Frontiers in Microbiology

    Article Title: Preparation of MS2 Phage-Like Particles and Their Use As Potential Process Control Viruses for Detection and Quantification of Enteric RNA Viruses in Different Matrices

    doi: 10.3389/fmicb.2016.01911

    Figure Lengend Snippet: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE – left part of the picture) and Western blot ( right part of the picture) results. 1,6, E. coli BL21 (DE3) negative control; 2,5, IPTG-non-induced culture; 3,4, IPTG-induced culture. 13 kDa MS2 coat protein was massively produced in the IPTG-induced culture. M, marker Spectra multicolor broad range protein ladder (Fermentas), the marker values are in kDa.

    Article Snippet: To verify the production of MS2 coat protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis was carried out using a phage MS2 Coat Protein polyclonal antibody (Merck Millipore, USA) as primary antibody and Goat Anti-Rabbit IgG, Fc specific fragment (Jackson ImmunoResearch, UK) as secondary antibody.

    Techniques: Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot, Negative Control, Produced, Marker