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Merck KGaA e coli bl21
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

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Related Articles

Clone Assay:

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: Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens
Article Snippet: .. E. coli BL21(DE3) (Merck Millipore, Darmstadt, Germany) was used as the expression host. pJET1.2 (Fermentas, Maryland, USA) and pET-29a(+) (Merck Millipore) were used as cloning and expression vectors, respectively. .. P. agarexedens BCRC 17346 was cultured at 30 °C in nutrient broth medium supplemented with 0.1% (w/v) urea and 1% (w/v) glucose.

Article Title: Direct induction of microtubule branching by microtubule nucleation factor SSNA1
Article Snippet: Protein preparation and purification The DNAs of CrSSNA1 and mouse SSNA1 were obtained by gene synthesis (GeneArt, ThermoFisher) and cloned into self-generated LIC (ligation-independent cloning) vectors. .. 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.

Amplification:

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).

Mass Spectrometry:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: Homogeneous human full-length protein was characterized by mass spectrometry for molecular weight determination and SDS-PAGE at reducing and non-reducing conditions followed by Coomassie staining, silver staining, or immunoblotting. .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

Construct:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck). ..

Incubation:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck). .. Purified TFPI-(1–150)-Thr was digested by incubation with thrombin resulting in the generation of TFPI-(1–83)-Thr and TFPI-(90–150)-Thr.

Expressing:

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: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck). ..

Article Title: Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens
Article Snippet: .. E. coli BL21(DE3) (Merck Millipore, Darmstadt, Germany) was used as the expression host. pJET1.2 (Fermentas, Maryland, USA) and pET-29a(+) (Merck Millipore) were used as cloning and expression vectors, respectively. .. P. agarexedens BCRC 17346 was cultured at 30 °C in nutrient broth medium supplemented with 0.1% (w/v) urea and 1% (w/v) glucose.

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: .. Expression of CV-1 to CV-5, CLC, IC-7, IL-6, and CNTF was performed in Escherichia coli BL21(DE3) (Merck KGaA). .. In detail, protein expression was induced at an A 600 of 0.8 with 1 m m isopropyl 1-thio-β- d -galactopyranoside.

Ligation:

Article Title: Direct induction of microtubule branching by microtubule nucleation factor SSNA1
Article Snippet: Protein preparation and purification The DNAs of CrSSNA1 and mouse SSNA1 were obtained by gene synthesis (GeneArt, ThermoFisher) and cloned into self-generated LIC (ligation-independent cloning) vectors. .. 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.

Cell Culture:

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: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: Cells were cultured in DMEM containing 10% fetal bovine serum (Life Technologies) and full-length TFPI was purified from conditioned media by a two-step affinity purification protocol. .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

Article Title: Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens
Article Snippet: E. coli BL21(DE3) (Merck Millipore, Darmstadt, Germany) was used as the expression host. pJET1.2 (Fermentas, Maryland, USA) and pET-29a(+) (Merck Millipore) were used as cloning and expression vectors, respectively. .. P. agarexedens BCRC 17346 was cultured at 30 °C in nutrient broth medium supplemented with 0.1% (w/v) urea and 1% (w/v) glucose.

Polymerase Chain Reaction:

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).

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.

Affinity Purification:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: Cells were cultured in DMEM containing 10% fetal bovine serum (Life Technologies) and full-length TFPI was purified from conditioned media by a two-step affinity purification protocol. .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

Recombinant:

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: Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens
Article Snippet: Escherichia coli ECOS™ 9-5 (Yeastern, Taipei, Taiwan) was used for the propagation and manipulation of recombinant DNA. .. E. coli BL21(DE3) (Merck Millipore, Darmstadt, Germany) was used as the expression host. pJET1.2 (Fermentas, Maryland, USA) and pET-29a(+) (Merck Millipore) were used as cloning and expression vectors, respectively.

Molecular Weight:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: Homogeneous human full-length protein was characterized by mass spectrometry for molecular weight determination and SDS-PAGE at reducing and non-reducing conditions followed by Coomassie staining, silver staining, or immunoblotting. .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

Isolation:

Article Title: Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens
Article Snippet: Bacterial strains, plasmids, and culture condition Paenibacillus agarexedens BCRC 17346 was purchased from BCRC (Bioresource Collection and Research Center, Hsinchu, Taiwan) and was used for the isolation of genomic DNA. .. E. coli BL21(DE3) (Merck Millipore, Darmstadt, Germany) was used as the expression host. pJET1.2 (Fermentas, Maryland, USA) and pET-29a(+) (Merck Millipore) were used as cloning and expression vectors, respectively.

Purification:

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: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: Paragraph title: Expression, Purification, and Characterization of TFPI Proteins ... The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

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: 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.

Positron Emission Tomography:

Article Title: Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens
Article Snippet: .. E. coli BL21(DE3) (Merck Millipore, Darmstadt, Germany) was used as the expression host. pJET1.2 (Fermentas, Maryland, USA) and pET-29a(+) (Merck Millipore) were used as cloning and expression vectors, respectively. .. P. agarexedens BCRC 17346 was cultured at 30 °C in nutrient broth medium supplemented with 0.1% (w/v) urea and 1% (w/v) glucose.

Affinity Column:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck). .. TFPI-(1-1–150)-Thr was purified using a two-step purification procedure, a Q-Sepharose column, and a streptavidin affinity column with an immobilized peptide specific toward TFPI.

Staining:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: Homogeneous human full-length protein was characterized by mass spectrometry for molecular weight determination and SDS-PAGE at reducing and non-reducing conditions followed by Coomassie staining, silver staining, or immunoblotting. .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

Silver Staining:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: Homogeneous human full-length protein was characterized by mass spectrometry for molecular weight determination and SDS-PAGE at reducing and non-reducing conditions followed by Coomassie staining, silver staining, or immunoblotting. .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

SDS Page:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: Homogeneous human full-length protein was characterized by mass spectrometry for molecular weight determination and SDS-PAGE at reducing and non-reducing conditions followed by Coomassie staining, silver staining, or immunoblotting. .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

Plasmid Preparation:

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: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck). ..

Affinity Chromatography:

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.

Concentration Assay:

Article Title: Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI) *
Article Snippet: The active concentration of full-length TFPI was determined by titrating a known amount of bovine FXa with TFPI ( ). .. The TFPI constructs TFPI-(1–150) and TFPI-(1–150)-Thr, containing a thrombin cleavage site between K1 and K2 were expressed as insoluble inclusion bodies in Escherichia coli BL21(DE3)pLysS (Merck, Darmstadt, Germany) using the expression vector pET19b (Merck).

Lysis:

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.

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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
    80/100 stars
      Buy from Supplier

    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
    83/100 stars
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    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

    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