Structured Review

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 15 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 15 article reviews
Price from $9.99 to $1999.99
e coli bl21 - by Bioz Stars, 2020-07
93/100 stars

Images

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

Related Articles

Clone Assay:

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: 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.

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).

Purification:

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.

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.

Polymerase Chain Reaction:

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.

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.

Expressing:

Article Title: Gene Cloning and Characterization of Two NADH-Dependent 3-Quinuclidinone Reductases from Microbacterium luteolum JCM 9174
Article Snippet: .. E. coli BL21(DE3) (Merck KGaA, Darmstadt, Germany) was used for the expression of pET28-QNR, and E. coli BL21(pET28-QNR) was cultivated at 37°C for 24 h in LB medium containing 0.05 mg ml−1 kanamycin sulfate and 0.4 mM isopropyl-β- d -1-thiogalactopyranoside (IPTG). .. The oligonucleotide primers of forward primer 1 (5′-ACNGCNYTNGTBACNGGYGGY-3′) and reverse primer 2 (5′-NCCNGTNAYRTARCTNGCNGCNGG-3′) were synthesized based on the conserved amino acid sequences of tropinone reductase I and II of Hyoscyamus niger (see ) , which are known to reduce 3-quinuclidinone.

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: 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: 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.

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.

Transformation Assay:

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 ( , ).

Recombinant:

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.

other:

Article Title: Genome-Wide Screening for Identification of Novel Toxin-Antitoxin Systems in Staphylococcus aureus
Article Snippet: E. coli BL21(DE3) strains carrying pBAD24 derivatives were grown with shaking at 37°C in 10 ml M9 Gly medium supplemented with ampicillin (100 μg/ml).

Plasmid Preparation:

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: 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 ( , ).

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 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 15 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 15 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    91
    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: 91/100, 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 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    bl21 de3 plyss strain - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    90
    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: 90/100, 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 90 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, 2020-07
    90/100 stars
      Buy from Supplier

    93
    Merck KGaA escherichia coli
    Expression, purification, and RNase activity of r Af RNASET2. (A) Analysis of the prokaryotic recombinant expression vector pET-His- Af RNASET2 (without signal peptides, Glutamic acid 26-Arginine 407) by double enzyme digestion. Lane M indicates the DNA marker. The 1,200 bp band indicates the Af RNASET2 cDNA insert. (B) SDS-PAGE analysis of pET-His-Af RNASET2 expression. Lane M, protein marker; lane 1, E. coli transformed with pET-His-Af RNASET2 prior to IPTG induction; lane 2, E. coli transformed with pET-His- Af RNASET2 following IPTG induction; lane 3, sediment from E. coli transformed with pET-His- Af RNASET2 plasmid following IPTG induction, and ultrasonication. (C) SDS-PAGE of renatured r Af RNASET2 protein following purification using Ni-NTA affinity chromatography. Lane M indicates the protein marker. (D) Analysis of RNase activity of r Af RNASET2 (1 µ g) with yeast RNA (50 µ g) at 37°C at 20, 40, 60, 80 and 100 min). (E) Analysis of RNase activity of r Af RNASET2 with or without RNasin (RNAse inhibitor) pretreatment. RNase enzymolysis products were analyzed using 1% agarose gel electrophoresis. E. coli, <t>Escherichia</t> coli ; r, recombinant; Af, Aspergillus fumigatus ; RNASET2, T2 ribonuclease; IPTG, isopropyl-β-d-thiogalactopyranoside.
    Escherichia Coli, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/escherichia coli/product/Merck KGaA
    Average 93 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    escherichia coli - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    Image Search Results


    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

    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

    Expression, purification, and RNase activity of r Af RNASET2. (A) Analysis of the prokaryotic recombinant expression vector pET-His- Af RNASET2 (without signal peptides, Glutamic acid 26-Arginine 407) by double enzyme digestion. Lane M indicates the DNA marker. The 1,200 bp band indicates the Af RNASET2 cDNA insert. (B) SDS-PAGE analysis of pET-His-Af RNASET2 expression. Lane M, protein marker; lane 1, E. coli transformed with pET-His-Af RNASET2 prior to IPTG induction; lane 2, E. coli transformed with pET-His- Af RNASET2 following IPTG induction; lane 3, sediment from E. coli transformed with pET-His- Af RNASET2 plasmid following IPTG induction, and ultrasonication. (C) SDS-PAGE of renatured r Af RNASET2 protein following purification using Ni-NTA affinity chromatography. Lane M indicates the protein marker. (D) Analysis of RNase activity of r Af RNASET2 (1 µ g) with yeast RNA (50 µ g) at 37°C at 20, 40, 60, 80 and 100 min). (E) Analysis of RNase activity of r Af RNASET2 with or without RNasin (RNAse inhibitor) pretreatment. RNase enzymolysis products were analyzed using 1% agarose gel electrophoresis. E. coli, Escherichia coli ; r, recombinant; Af, Aspergillus fumigatus ; RNASET2, T2 ribonuclease; IPTG, isopropyl-β-d-thiogalactopyranoside.

    Journal: International Journal of Molecular Medicine

    Article Title: Ribonuclease T2 from Aspergillus fumigatus promotes T helper type 2 responses through M2 polarization of macrophages

    doi: 10.3892/ijmm.2020.4613

    Figure Lengend Snippet: Expression, purification, and RNase activity of r Af RNASET2. (A) Analysis of the prokaryotic recombinant expression vector pET-His- Af RNASET2 (without signal peptides, Glutamic acid 26-Arginine 407) by double enzyme digestion. Lane M indicates the DNA marker. The 1,200 bp band indicates the Af RNASET2 cDNA insert. (B) SDS-PAGE analysis of pET-His-Af RNASET2 expression. Lane M, protein marker; lane 1, E. coli transformed with pET-His-Af RNASET2 prior to IPTG induction; lane 2, E. coli transformed with pET-His- Af RNASET2 following IPTG induction; lane 3, sediment from E. coli transformed with pET-His- Af RNASET2 plasmid following IPTG induction, and ultrasonication. (C) SDS-PAGE of renatured r Af RNASET2 protein following purification using Ni-NTA affinity chromatography. Lane M indicates the protein marker. (D) Analysis of RNase activity of r Af RNASET2 (1 µ g) with yeast RNA (50 µ g) at 37°C at 20, 40, 60, 80 and 100 min). (E) Analysis of RNase activity of r Af RNASET2 with or without RNasin (RNAse inhibitor) pretreatment. RNase enzymolysis products were analyzed using 1% agarose gel electrophoresis. E. coli, Escherichia coli ; r, recombinant; Af, Aspergillus fumigatus ; RNASET2, T2 ribonuclease; IPTG, isopropyl-β-d-thiogalactopyranoside.

    Article Snippet: Expression system components and reagents The following reagents were purchased for recombinant protein expression: Escherichia coli (E. coli ) BL21 (DE3) plysS cells (Merck KGaA), pMD 19-T vectors (Takara Biotechnology Co., Ltd.), pET-His vectors (Wuhan Miaoling Bioscience & Technology Co., Ltd.), and primer STAR HS DNA polymerase (Takara Biotechnology Co., Ltd.).

    Techniques: Expressing, Purification, Activity Assay, Recombinant, Plasmid Preparation, Positron Emission Tomography, Marker, SDS Page, Transformation Assay, Affinity Chromatography, Agarose Gel Electrophoresis