5 ml histrap ff crude column  (GE Healthcare)

 
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    HisTrap FF
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    Structured Review

    GE Healthcare 5 ml histrap ff crude column

    https://www.bioz.com/result/5 ml histrap ff crude column/product/GE Healthcare
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    5 ml histrap ff crude column - by Bioz Stars, 2021-03
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    Purification:

    Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
    Article Snippet: At that time, the recombinant E. coli was induced by adding 1 mM isopropyl-β- d -thio-galactoside (IPTG) and was cultured for another 5 h. The cells were then harvested by centrifugation, washed twice with phosphate-buffered saline (PBS), and resuspended in 8 ml of 50 mM sodium phosphate (pH 7.5) containing 500 mM NaCl and 20 mM imidazole. .. The supernatant was saved for subsequent purification of the 6×His-tagged fusion proteins using a HisTrap FF column (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) according to the manufacturer's instructions. ..

    Article Title: Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification
    Article Snippet: Cells were lysed by sonication and centrifuged to pellet cell debris as above. .. The soluble supernatant fraction was purified on a 5 ml HisTrap ff crude column (GE Healthcare, 17-5286-01) using the AKTA protein purification system (GE Healthcare). .. The column was washed with ten column volumes of 20 mM HEPES pH7, 200 mM NaCl and 25 mM imidazole (Sigma Aldrich, I5513).

    Article Title: Processivity and enzymatic mechanism of a multifunctional family 5 endoglucanase from Bacillus subtilis BS-5 with potential applications in the saccharification of cellulosic substrates
    Article Snippet: Expression and purification of enzymes Recombinant E. coli BL21 cells harboring the plasmids described above were cultured in LB medium and the target genes were expressed as described above [ , ]. .. Recombinant proteins were purified by affinity chromatography with a HisTrap FF column (GE Healthcare, Fairfield, USA) according to the manufacturer’s recommendations. .. Enzyme homogeneity and the molecular weights of purified EG5C, EG5C-1 and EG5C-2 were estimated on a 12% SDS-PAGE gel.

    Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
    Article Snippet: To facilitate the high-level inducible expression of the cloned fusion gene, plasmid pET32a:: atl was transformed into E. coli BL21. .. The Atl fusion protein was extracted from a mid-log culture, which had been induced with 1 mM IPTG for 5 h. The IPTG-inducible expression plasmid pET32a contained a start codon followed by a 6×His tag sequence, allowing purification of the recombinant autolysin by using a HisTrap FF column. .. As shown in , an apparent protein band of approximately 96 kDa appeared in the induced recombinant E. coli , while it was absent in the uninduced recombinant E. coli or induced E. coli containing control plasmid.

    Centrifugation:

    Article Title: Engineering of monobody conjugates for human EphA2-specific optical imaging
    Article Snippet: The pellet was resuspended in 30 mL of Dulbecco’s phosphate buffered saline (DPBS) and lysed by sonication for 30 min at 4°C. .. Supernatants were obtained by centrifugation at 10,000 rpm at 4°C and applied to a HisTrap FF column (GE Healthcare Biosciences, PA) in an AKTA FPLC system (GE Healthcare Biosciences). ..

    Fast Protein Liquid Chromatography:

    Article Title: Engineering of monobody conjugates for human EphA2-specific optical imaging
    Article Snippet: The pellet was resuspended in 30 mL of Dulbecco’s phosphate buffered saline (DPBS) and lysed by sonication for 30 min at 4°C. .. Supernatants were obtained by centrifugation at 10,000 rpm at 4°C and applied to a HisTrap FF column (GE Healthcare Biosciences, PA) in an AKTA FPLC system (GE Healthcare Biosciences). ..

    Protein Purification:

    Article Title: Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification
    Article Snippet: Cells were lysed by sonication and centrifuged to pellet cell debris as above. .. The soluble supernatant fraction was purified on a 5 ml HisTrap ff crude column (GE Healthcare, 17-5286-01) using the AKTA protein purification system (GE Healthcare). .. The column was washed with ten column volumes of 20 mM HEPES pH7, 200 mM NaCl and 25 mM imidazole (Sigma Aldrich, I5513).

    Recombinant:

    Article Title: Processivity and enzymatic mechanism of a multifunctional family 5 endoglucanase from Bacillus subtilis BS-5 with potential applications in the saccharification of cellulosic substrates
    Article Snippet: Expression and purification of enzymes Recombinant E. coli BL21 cells harboring the plasmids described above were cultured in LB medium and the target genes were expressed as described above [ , ]. .. Recombinant proteins were purified by affinity chromatography with a HisTrap FF column (GE Healthcare, Fairfield, USA) according to the manufacturer’s recommendations. .. Enzyme homogeneity and the molecular weights of purified EG5C, EG5C-1 and EG5C-2 were estimated on a 12% SDS-PAGE gel.

    Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
    Article Snippet: To facilitate the high-level inducible expression of the cloned fusion gene, plasmid pET32a:: atl was transformed into E. coli BL21. .. The Atl fusion protein was extracted from a mid-log culture, which had been induced with 1 mM IPTG for 5 h. The IPTG-inducible expression plasmid pET32a contained a start codon followed by a 6×His tag sequence, allowing purification of the recombinant autolysin by using a HisTrap FF column. .. As shown in , an apparent protein band of approximately 96 kDa appeared in the induced recombinant E. coli , while it was absent in the uninduced recombinant E. coli or induced E. coli containing control plasmid.

    Affinity Chromatography:

    Article Title: Processivity and enzymatic mechanism of a multifunctional family 5 endoglucanase from Bacillus subtilis BS-5 with potential applications in the saccharification of cellulosic substrates
    Article Snippet: Expression and purification of enzymes Recombinant E. coli BL21 cells harboring the plasmids described above were cultured in LB medium and the target genes were expressed as described above [ , ]. .. Recombinant proteins were purified by affinity chromatography with a HisTrap FF column (GE Healthcare, Fairfield, USA) according to the manufacturer’s recommendations. .. Enzyme homogeneity and the molecular weights of purified EG5C, EG5C-1 and EG5C-2 were estimated on a 12% SDS-PAGE gel.

    Flow Cytometry:

    Article Title: Computational redesign of the Escherichia coli ribose-binding protein ligand binding pocket for 1,3-cyclohexanediol and cyclohexanol
    Article Snippet: .. The clean lysate was next loaded onto a HisTrap HP column (HisTrap FF crude 1 ml, GE Healthcare) at 4 °C and flow rate of 0.5 ml min−1 , followed by washes of, consecutively, 10 column volumes (cv, equal to 1 ml) of buffer A with 20 mM imidazole, 1.5 cv of buffer A with 40 mM imidazole and 1.5 cv of buffer A with 80 mM imidazole. ..

    Expressing:

    Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
    Article Snippet: To facilitate the high-level inducible expression of the cloned fusion gene, plasmid pET32a:: atl was transformed into E. coli BL21. .. The Atl fusion protein was extracted from a mid-log culture, which had been induced with 1 mM IPTG for 5 h. The IPTG-inducible expression plasmid pET32a contained a start codon followed by a 6×His tag sequence, allowing purification of the recombinant autolysin by using a HisTrap FF column. .. As shown in , an apparent protein band of approximately 96 kDa appeared in the induced recombinant E. coli , while it was absent in the uninduced recombinant E. coli or induced E. coli containing control plasmid.

    Plasmid Preparation:

    Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
    Article Snippet: To facilitate the high-level inducible expression of the cloned fusion gene, plasmid pET32a:: atl was transformed into E. coli BL21. .. The Atl fusion protein was extracted from a mid-log culture, which had been induced with 1 mM IPTG for 5 h. The IPTG-inducible expression plasmid pET32a contained a start codon followed by a 6×His tag sequence, allowing purification of the recombinant autolysin by using a HisTrap FF column. .. As shown in , an apparent protein band of approximately 96 kDa appeared in the induced recombinant E. coli , while it was absent in the uninduced recombinant E. coli or induced E. coli containing control plasmid.

    Sequencing:

    Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
    Article Snippet: To facilitate the high-level inducible expression of the cloned fusion gene, plasmid pET32a:: atl was transformed into E. coli BL21. .. The Atl fusion protein was extracted from a mid-log culture, which had been induced with 1 mM IPTG for 5 h. The IPTG-inducible expression plasmid pET32a contained a start codon followed by a 6×His tag sequence, allowing purification of the recombinant autolysin by using a HisTrap FF column. .. As shown in , an apparent protein band of approximately 96 kDa appeared in the induced recombinant E. coli , while it was absent in the uninduced recombinant E. coli or induced E. coli containing control plasmid.

    Lysis:

    Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
    Article Snippet: These purified proteins are a useful in vitro model for the characterization of the normal and mutant human tyrosinase structure and function and, potentially, the discovery of novel therapeutics. .. T.ni larvae biomass containing the protein of interest (Allotropic Tech., MD) Wet ice 50 ml tubes (Corning, NY) Lysis buffer (see recipe) 1-Phenyl-2-thiourea (PTU; Sigma-Aldrich, MO) DNase I (Thermo Fisher Scientific, PA) Complete set of protease inhibitors (Roche, USA) tris(2-carboxyethyl)phosphine (TCEP; Thermo Scientific, DE) Fisher Science Education pH meter (Thermo Fisher Scientific, MA) Omni Tissue Homogenizer TH (Omni International, GA) Hard Tissue Omni Tip™ Homogenizer Probes (Omni International, GA) Orbitron Rotator (Boekel Scientific, PA) Ultrasonic Processor GE130PB (Hielscher System, Germany) Sorvall Lynx 4000 Centrifuge (Thermo Scientific, DE) Rotor for centrifuge: Fiberlite, F21-8×50y (capacity 8 × 50 ml) Binding buffer (see recipe) Elution buffer (see recipe) Immobilized metal affinity (IMAC) column - HisTrap FF Crude (GE Healthcare, NJ) BioLogic Duo-Flow chromatography system (Bio-Rad, CA) Gel-filtration buffer (see recipe) NanoDrop 2000c UV-Vis spectrophotometer (Thermo Scientific, DE) 4-15% polyacrylamide gels (Bio-Rad, CA) 3,4-Dihydroxy-L-phenylalanine, L-DOPA (Sigma Aldrich, MO) MO) SpectraMax i3 Multi-Mode Microplate Detection Platform (Molecular Devices, CA) Clear-bottom polystyrene 96-well plates (Corning, NY) Dialysis membranes: SnakeSkin dialysis tubing or Slide-A-Lyzer dialysis cassettes, 10K MWCO (ThermoFisher Scientific, MA) Gel-filtration columns: HiPrep 16/60 Sephacryl S-100 and Superdex 75 10/300 (GE HealthCare, NJ) Protein standards (Bio-Rad, CA) Anti-His antibody (Life Technologies, NY) Anti-tyrosinase antibody (T311 Santa Cruz Biotechnology, CA) Centrifuge 5415R (Eppendorf, Germany) UN-SCAN-IT gel ™ gel analysis software (Silk Scientific, Inc., UT) Stirring devices Centrifuge tubes Additional materials and equipment for SDS-PAGE ( UNIT 10.1 ) and Western blot ( UNIT 10.8 ) analysis. .. The DNA of hTyrCtr included residues 19-469 of the human tyrosinase, a TEV-protease cleavage site (NLYFQG sequence), and a 6×His-Tag ( ) was commercially synthesized, expressed in baculovirus and produced in whole insect T. ni larvae (Allotropic Tech., MD).

    Binding Assay:

    Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
    Article Snippet: These purified proteins are a useful in vitro model for the characterization of the normal and mutant human tyrosinase structure and function and, potentially, the discovery of novel therapeutics. .. T.ni larvae biomass containing the protein of interest (Allotropic Tech., MD) Wet ice 50 ml tubes (Corning, NY) Lysis buffer (see recipe) 1-Phenyl-2-thiourea (PTU; Sigma-Aldrich, MO) DNase I (Thermo Fisher Scientific, PA) Complete set of protease inhibitors (Roche, USA) tris(2-carboxyethyl)phosphine (TCEP; Thermo Scientific, DE) Fisher Science Education pH meter (Thermo Fisher Scientific, MA) Omni Tissue Homogenizer TH (Omni International, GA) Hard Tissue Omni Tip™ Homogenizer Probes (Omni International, GA) Orbitron Rotator (Boekel Scientific, PA) Ultrasonic Processor GE130PB (Hielscher System, Germany) Sorvall Lynx 4000 Centrifuge (Thermo Scientific, DE) Rotor for centrifuge: Fiberlite, F21-8×50y (capacity 8 × 50 ml) Binding buffer (see recipe) Elution buffer (see recipe) Immobilized metal affinity (IMAC) column - HisTrap FF Crude (GE Healthcare, NJ) BioLogic Duo-Flow chromatography system (Bio-Rad, CA) Gel-filtration buffer (see recipe) NanoDrop 2000c UV-Vis spectrophotometer (Thermo Scientific, DE) 4-15% polyacrylamide gels (Bio-Rad, CA) 3,4-Dihydroxy-L-phenylalanine, L-DOPA (Sigma Aldrich, MO) MO) SpectraMax i3 Multi-Mode Microplate Detection Platform (Molecular Devices, CA) Clear-bottom polystyrene 96-well plates (Corning, NY) Dialysis membranes: SnakeSkin dialysis tubing or Slide-A-Lyzer dialysis cassettes, 10K MWCO (ThermoFisher Scientific, MA) Gel-filtration columns: HiPrep 16/60 Sephacryl S-100 and Superdex 75 10/300 (GE HealthCare, NJ) Protein standards (Bio-Rad, CA) Anti-His antibody (Life Technologies, NY) Anti-tyrosinase antibody (T311 Santa Cruz Biotechnology, CA) Centrifuge 5415R (Eppendorf, Germany) UN-SCAN-IT gel ™ gel analysis software (Silk Scientific, Inc., UT) Stirring devices Centrifuge tubes Additional materials and equipment for SDS-PAGE ( UNIT 10.1 ) and Western blot ( UNIT 10.8 ) analysis. .. The DNA of hTyrCtr included residues 19-469 of the human tyrosinase, a TEV-protease cleavage site (NLYFQG sequence), and a 6×His-Tag ( ) was commercially synthesized, expressed in baculovirus and produced in whole insect T. ni larvae (Allotropic Tech., MD).

    Chromatography:

    Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
    Article Snippet: These purified proteins are a useful in vitro model for the characterization of the normal and mutant human tyrosinase structure and function and, potentially, the discovery of novel therapeutics. .. T.ni larvae biomass containing the protein of interest (Allotropic Tech., MD) Wet ice 50 ml tubes (Corning, NY) Lysis buffer (see recipe) 1-Phenyl-2-thiourea (PTU; Sigma-Aldrich, MO) DNase I (Thermo Fisher Scientific, PA) Complete set of protease inhibitors (Roche, USA) tris(2-carboxyethyl)phosphine (TCEP; Thermo Scientific, DE) Fisher Science Education pH meter (Thermo Fisher Scientific, MA) Omni Tissue Homogenizer TH (Omni International, GA) Hard Tissue Omni Tip™ Homogenizer Probes (Omni International, GA) Orbitron Rotator (Boekel Scientific, PA) Ultrasonic Processor GE130PB (Hielscher System, Germany) Sorvall Lynx 4000 Centrifuge (Thermo Scientific, DE) Rotor for centrifuge: Fiberlite, F21-8×50y (capacity 8 × 50 ml) Binding buffer (see recipe) Elution buffer (see recipe) Immobilized metal affinity (IMAC) column - HisTrap FF Crude (GE Healthcare, NJ) BioLogic Duo-Flow chromatography system (Bio-Rad, CA) Gel-filtration buffer (see recipe) NanoDrop 2000c UV-Vis spectrophotometer (Thermo Scientific, DE) 4-15% polyacrylamide gels (Bio-Rad, CA) 3,4-Dihydroxy-L-phenylalanine, L-DOPA (Sigma Aldrich, MO) MO) SpectraMax i3 Multi-Mode Microplate Detection Platform (Molecular Devices, CA) Clear-bottom polystyrene 96-well plates (Corning, NY) Dialysis membranes: SnakeSkin dialysis tubing or Slide-A-Lyzer dialysis cassettes, 10K MWCO (ThermoFisher Scientific, MA) Gel-filtration columns: HiPrep 16/60 Sephacryl S-100 and Superdex 75 10/300 (GE HealthCare, NJ) Protein standards (Bio-Rad, CA) Anti-His antibody (Life Technologies, NY) Anti-tyrosinase antibody (T311 Santa Cruz Biotechnology, CA) Centrifuge 5415R (Eppendorf, Germany) UN-SCAN-IT gel ™ gel analysis software (Silk Scientific, Inc., UT) Stirring devices Centrifuge tubes Additional materials and equipment for SDS-PAGE ( UNIT 10.1 ) and Western blot ( UNIT 10.8 ) analysis. .. The DNA of hTyrCtr included residues 19-469 of the human tyrosinase, a TEV-protease cleavage site (NLYFQG sequence), and a 6×His-Tag ( ) was commercially synthesized, expressed in baculovirus and produced in whole insect T. ni larvae (Allotropic Tech., MD).

    Spectrophotometry:

    Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
    Article Snippet: These purified proteins are a useful in vitro model for the characterization of the normal and mutant human tyrosinase structure and function and, potentially, the discovery of novel therapeutics. .. T.ni larvae biomass containing the protein of interest (Allotropic Tech., MD) Wet ice 50 ml tubes (Corning, NY) Lysis buffer (see recipe) 1-Phenyl-2-thiourea (PTU; Sigma-Aldrich, MO) DNase I (Thermo Fisher Scientific, PA) Complete set of protease inhibitors (Roche, USA) tris(2-carboxyethyl)phosphine (TCEP; Thermo Scientific, DE) Fisher Science Education pH meter (Thermo Fisher Scientific, MA) Omni Tissue Homogenizer TH (Omni International, GA) Hard Tissue Omni Tip™ Homogenizer Probes (Omni International, GA) Orbitron Rotator (Boekel Scientific, PA) Ultrasonic Processor GE130PB (Hielscher System, Germany) Sorvall Lynx 4000 Centrifuge (Thermo Scientific, DE) Rotor for centrifuge: Fiberlite, F21-8×50y (capacity 8 × 50 ml) Binding buffer (see recipe) Elution buffer (see recipe) Immobilized metal affinity (IMAC) column - HisTrap FF Crude (GE Healthcare, NJ) BioLogic Duo-Flow chromatography system (Bio-Rad, CA) Gel-filtration buffer (see recipe) NanoDrop 2000c UV-Vis spectrophotometer (Thermo Scientific, DE) 4-15% polyacrylamide gels (Bio-Rad, CA) 3,4-Dihydroxy-L-phenylalanine, L-DOPA (Sigma Aldrich, MO) MO) SpectraMax i3 Multi-Mode Microplate Detection Platform (Molecular Devices, CA) Clear-bottom polystyrene 96-well plates (Corning, NY) Dialysis membranes: SnakeSkin dialysis tubing or Slide-A-Lyzer dialysis cassettes, 10K MWCO (ThermoFisher Scientific, MA) Gel-filtration columns: HiPrep 16/60 Sephacryl S-100 and Superdex 75 10/300 (GE HealthCare, NJ) Protein standards (Bio-Rad, CA) Anti-His antibody (Life Technologies, NY) Anti-tyrosinase antibody (T311 Santa Cruz Biotechnology, CA) Centrifuge 5415R (Eppendorf, Germany) UN-SCAN-IT gel ™ gel analysis software (Silk Scientific, Inc., UT) Stirring devices Centrifuge tubes Additional materials and equipment for SDS-PAGE ( UNIT 10.1 ) and Western blot ( UNIT 10.8 ) analysis. .. The DNA of hTyrCtr included residues 19-469 of the human tyrosinase, a TEV-protease cleavage site (NLYFQG sequence), and a 6×His-Tag ( ) was commercially synthesized, expressed in baculovirus and produced in whole insect T. ni larvae (Allotropic Tech., MD).

    Software:

    Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
    Article Snippet: These purified proteins are a useful in vitro model for the characterization of the normal and mutant human tyrosinase structure and function and, potentially, the discovery of novel therapeutics. .. T.ni larvae biomass containing the protein of interest (Allotropic Tech., MD) Wet ice 50 ml tubes (Corning, NY) Lysis buffer (see recipe) 1-Phenyl-2-thiourea (PTU; Sigma-Aldrich, MO) DNase I (Thermo Fisher Scientific, PA) Complete set of protease inhibitors (Roche, USA) tris(2-carboxyethyl)phosphine (TCEP; Thermo Scientific, DE) Fisher Science Education pH meter (Thermo Fisher Scientific, MA) Omni Tissue Homogenizer TH (Omni International, GA) Hard Tissue Omni Tip™ Homogenizer Probes (Omni International, GA) Orbitron Rotator (Boekel Scientific, PA) Ultrasonic Processor GE130PB (Hielscher System, Germany) Sorvall Lynx 4000 Centrifuge (Thermo Scientific, DE) Rotor for centrifuge: Fiberlite, F21-8×50y (capacity 8 × 50 ml) Binding buffer (see recipe) Elution buffer (see recipe) Immobilized metal affinity (IMAC) column - HisTrap FF Crude (GE Healthcare, NJ) BioLogic Duo-Flow chromatography system (Bio-Rad, CA) Gel-filtration buffer (see recipe) NanoDrop 2000c UV-Vis spectrophotometer (Thermo Scientific, DE) 4-15% polyacrylamide gels (Bio-Rad, CA) 3,4-Dihydroxy-L-phenylalanine, L-DOPA (Sigma Aldrich, MO) MO) SpectraMax i3 Multi-Mode Microplate Detection Platform (Molecular Devices, CA) Clear-bottom polystyrene 96-well plates (Corning, NY) Dialysis membranes: SnakeSkin dialysis tubing or Slide-A-Lyzer dialysis cassettes, 10K MWCO (ThermoFisher Scientific, MA) Gel-filtration columns: HiPrep 16/60 Sephacryl S-100 and Superdex 75 10/300 (GE HealthCare, NJ) Protein standards (Bio-Rad, CA) Anti-His antibody (Life Technologies, NY) Anti-tyrosinase antibody (T311 Santa Cruz Biotechnology, CA) Centrifuge 5415R (Eppendorf, Germany) UN-SCAN-IT gel ™ gel analysis software (Silk Scientific, Inc., UT) Stirring devices Centrifuge tubes Additional materials and equipment for SDS-PAGE ( UNIT 10.1 ) and Western blot ( UNIT 10.8 ) analysis. .. The DNA of hTyrCtr included residues 19-469 of the human tyrosinase, a TEV-protease cleavage site (NLYFQG sequence), and a 6×His-Tag ( ) was commercially synthesized, expressed in baculovirus and produced in whole insect T. ni larvae (Allotropic Tech., MD).

    SDS Page:

    Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
    Article Snippet: These purified proteins are a useful in vitro model for the characterization of the normal and mutant human tyrosinase structure and function and, potentially, the discovery of novel therapeutics. .. T.ni larvae biomass containing the protein of interest (Allotropic Tech., MD) Wet ice 50 ml tubes (Corning, NY) Lysis buffer (see recipe) 1-Phenyl-2-thiourea (PTU; Sigma-Aldrich, MO) DNase I (Thermo Fisher Scientific, PA) Complete set of protease inhibitors (Roche, USA) tris(2-carboxyethyl)phosphine (TCEP; Thermo Scientific, DE) Fisher Science Education pH meter (Thermo Fisher Scientific, MA) Omni Tissue Homogenizer TH (Omni International, GA) Hard Tissue Omni Tip™ Homogenizer Probes (Omni International, GA) Orbitron Rotator (Boekel Scientific, PA) Ultrasonic Processor GE130PB (Hielscher System, Germany) Sorvall Lynx 4000 Centrifuge (Thermo Scientific, DE) Rotor for centrifuge: Fiberlite, F21-8×50y (capacity 8 × 50 ml) Binding buffer (see recipe) Elution buffer (see recipe) Immobilized metal affinity (IMAC) column - HisTrap FF Crude (GE Healthcare, NJ) BioLogic Duo-Flow chromatography system (Bio-Rad, CA) Gel-filtration buffer (see recipe) NanoDrop 2000c UV-Vis spectrophotometer (Thermo Scientific, DE) 4-15% polyacrylamide gels (Bio-Rad, CA) 3,4-Dihydroxy-L-phenylalanine, L-DOPA (Sigma Aldrich, MO) MO) SpectraMax i3 Multi-Mode Microplate Detection Platform (Molecular Devices, CA) Clear-bottom polystyrene 96-well plates (Corning, NY) Dialysis membranes: SnakeSkin dialysis tubing or Slide-A-Lyzer dialysis cassettes, 10K MWCO (ThermoFisher Scientific, MA) Gel-filtration columns: HiPrep 16/60 Sephacryl S-100 and Superdex 75 10/300 (GE HealthCare, NJ) Protein standards (Bio-Rad, CA) Anti-His antibody (Life Technologies, NY) Anti-tyrosinase antibody (T311 Santa Cruz Biotechnology, CA) Centrifuge 5415R (Eppendorf, Germany) UN-SCAN-IT gel ™ gel analysis software (Silk Scientific, Inc., UT) Stirring devices Centrifuge tubes Additional materials and equipment for SDS-PAGE ( UNIT 10.1 ) and Western blot ( UNIT 10.8 ) analysis. .. The DNA of hTyrCtr included residues 19-469 of the human tyrosinase, a TEV-protease cleavage site (NLYFQG sequence), and a 6×His-Tag ( ) was commercially synthesized, expressed in baculovirus and produced in whole insect T. ni larvae (Allotropic Tech., MD).

    Western Blot:

    Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
    Article Snippet: These purified proteins are a useful in vitro model for the characterization of the normal and mutant human tyrosinase structure and function and, potentially, the discovery of novel therapeutics. .. T.ni larvae biomass containing the protein of interest (Allotropic Tech., MD) Wet ice 50 ml tubes (Corning, NY) Lysis buffer (see recipe) 1-Phenyl-2-thiourea (PTU; Sigma-Aldrich, MO) DNase I (Thermo Fisher Scientific, PA) Complete set of protease inhibitors (Roche, USA) tris(2-carboxyethyl)phosphine (TCEP; Thermo Scientific, DE) Fisher Science Education pH meter (Thermo Fisher Scientific, MA) Omni Tissue Homogenizer TH (Omni International, GA) Hard Tissue Omni Tip™ Homogenizer Probes (Omni International, GA) Orbitron Rotator (Boekel Scientific, PA) Ultrasonic Processor GE130PB (Hielscher System, Germany) Sorvall Lynx 4000 Centrifuge (Thermo Scientific, DE) Rotor for centrifuge: Fiberlite, F21-8×50y (capacity 8 × 50 ml) Binding buffer (see recipe) Elution buffer (see recipe) Immobilized metal affinity (IMAC) column - HisTrap FF Crude (GE Healthcare, NJ) BioLogic Duo-Flow chromatography system (Bio-Rad, CA) Gel-filtration buffer (see recipe) NanoDrop 2000c UV-Vis spectrophotometer (Thermo Scientific, DE) 4-15% polyacrylamide gels (Bio-Rad, CA) 3,4-Dihydroxy-L-phenylalanine, L-DOPA (Sigma Aldrich, MO) MO) SpectraMax i3 Multi-Mode Microplate Detection Platform (Molecular Devices, CA) Clear-bottom polystyrene 96-well plates (Corning, NY) Dialysis membranes: SnakeSkin dialysis tubing or Slide-A-Lyzer dialysis cassettes, 10K MWCO (ThermoFisher Scientific, MA) Gel-filtration columns: HiPrep 16/60 Sephacryl S-100 and Superdex 75 10/300 (GE HealthCare, NJ) Protein standards (Bio-Rad, CA) Anti-His antibody (Life Technologies, NY) Anti-tyrosinase antibody (T311 Santa Cruz Biotechnology, CA) Centrifuge 5415R (Eppendorf, Germany) UN-SCAN-IT gel ™ gel analysis software (Silk Scientific, Inc., UT) Stirring devices Centrifuge tubes Additional materials and equipment for SDS-PAGE ( UNIT 10.1 ) and Western blot ( UNIT 10.8 ) analysis. .. The DNA of hTyrCtr included residues 19-469 of the human tyrosinase, a TEV-protease cleavage site (NLYFQG sequence), and a 6×His-Tag ( ) was commercially synthesized, expressed in baculovirus and produced in whole insect T. ni larvae (Allotropic Tech., MD).

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    GE Healthcare histrap ff crude column
    Purified trKMO. (A) Chromatogram produced during NiNTA affinity purification of truncate KMO. (B) SDS-PAGE protein gel stained with simplyblue showing <t>HisTrap</t> purification of truncated human KMO protein, (a) Cell-free extract prior to affinity chromatography, (b) flowthrough proteins which did not bind to the column, (c) first column wash, (d) second column wash, (e) third column wash, (f) pure truncated human KMO protein (44 kDa) from the fractions under the chromatogram. (C) Cartoon illustrating the trKMO construct. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
    Histrap Ff Crude Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histrap ff crude column/product/GE Healthcare
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    Price from $9.99 to $1999.99
    histrap ff crude column - by Bioz Stars, 2021-03
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    Purified trKMO. (A) Chromatogram produced during NiNTA affinity purification of truncate KMO. (B) SDS-PAGE protein gel stained with simplyblue showing HisTrap purification of truncated human KMO protein, (a) Cell-free extract prior to affinity chromatography, (b) flowthrough proteins which did not bind to the column, (c) first column wash, (d) second column wash, (e) third column wash, (f) pure truncated human KMO protein (44 kDa) from the fractions under the chromatogram. (C) Cartoon illustrating the trKMO construct. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: Protein Expression and Purification

    Article Title: Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification

    doi: 10.1016/j.pep.2013.11.015

    Figure Lengend Snippet: Purified trKMO. (A) Chromatogram produced during NiNTA affinity purification of truncate KMO. (B) SDS-PAGE protein gel stained with simplyblue showing HisTrap purification of truncated human KMO protein, (a) Cell-free extract prior to affinity chromatography, (b) flowthrough proteins which did not bind to the column, (c) first column wash, (d) second column wash, (e) third column wash, (f) pure truncated human KMO protein (44 kDa) from the fractions under the chromatogram. (C) Cartoon illustrating the trKMO construct. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: The soluble supernatant fraction was purified on a 5 ml HisTrap ff crude column (GE Healthcare, 17-5286-01) using the AKTA protein purification system (GE Healthcare).

    Techniques: Purification, Produced, Affinity Purification, SDS Page, Staining, Affinity Chromatography, Construct