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GE Healthcare histrap ff crude chromatography columns
Histrap Ff Crude Chromatography Columns, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/histrap ff crude chromatography columns/product/GE Healthcare
Average 99 stars, based on 1 article reviews
Price from $9.99 to $1999.99
histrap ff crude chromatography columns - by Bioz Stars, 2020-09
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Centrifugation:

Article Title: Engineering of monobody conjugates for human EphA2-specific optical imaging
Article Snippet: .. 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). ..

Recombinant:

Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
Article Snippet: .. 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.

Chromatography:

Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
Article Snippet: .. 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).

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. ..

Spectrophotometry:

Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
Article Snippet: .. 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).

Purification:

Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
Article Snippet: .. 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: .. 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: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
Article Snippet: .. 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.

Article Title: Large-scale seroprevalence analysis of human metapneumovirus and human respiratory syncytial virus infections in Beijing, China
Article Snippet: .. The supernatant was purified using a HisTrap Q FF 5 ml column (GE Healthcare) followed by a HisTrap HP 1 ml column (GE Healthcare). .. The purification of N proteins was evaluated by 12% SDS-PAGE and optical density scanning using the Gene Genius Bio-imaging System (Syngene, Synoptics Ltd. Cambridge.

Protein Purification:

Article Title: Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification
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). .. The column was washed with ten column volumes of 20 mM HEPES pH7, 200 mM NaCl and 25 mM imidazole (Sigma Aldrich, I5513).

Fast Protein Liquid Chromatography:

Article Title: Engineering of monobody conjugates for human EphA2-specific optical imaging
Article Snippet: .. 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). ..

Expressing:

Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
Article Snippet: .. 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: .. 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).

Sequencing:

Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
Article Snippet: .. 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.

Western Blot:

Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
Article Snippet: .. 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: .. 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: .. 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).

Plasmid Preparation:

Article Title: Characterization and Functional Analysis of atl, a Novel Gene Encoding Autolysin in Streptococcus suis
Article Snippet: .. 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.

Software:

Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector
Article Snippet: .. 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 metal affinity imac column histrap ff crude
    Chromatography profiles and enzymatic activities for the recombinant hTyrC tr and temperature sensitive mutant variant, R422Q A: <t>IMAC</t> using <t>HisTrap</t> FF crude 5 ml column with the linear gradient of 500 mM imidazole (gray line). B: Gel-filtration chromatography using HiPrep 16/60 Sephacryl S-100 column. The insert shows a magnification of A 280 profiles of hTyrC tr (black line) and R422Q mutant (green line). C: Gel filtration using Superdex 75 10/30 column. Top panels of A, B, and C show test tubes containing the L-DOPA colorimetric reactions for each protein fraction of hTyrC tr (black frame) and R422Q (green frame). Brown color (intensity proportional) in tube indicates diphenol oxidase activity.
    Metal Affinity Imac Column Histrap Ff Crude, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 257 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/metal affinity imac column histrap ff crude/product/GE Healthcare
    Average 99 stars, based on 257 article reviews
    Price from $9.99 to $1999.99
    metal affinity imac column histrap ff crude - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    GE Healthcare ge healthcare histrap crude
    Recombinant hTyr purified from T . ni . larval biomass in presence of hydrogenated Triton X-100. Panel A : hTyr eluted from Superdex 200 increase 10/300 GL column after Ni-NTA gravity purification. The absorption was measured at 280 nm (black solid line) and 260 nm (black dashed line). The following Bio-Rad SEC standards are shown at the top of the panel: Thyroglobulin (670 kDa), γ-globulin (158 kDa), and ovalbumin (44 kDa). The insert shows SDS-PAGE ( a ), Western blot ( b ), and diphenol oxidase activity ( c ) for the corresponding fractions containing hTyr. Arrows display the protein ladder marker at 70 kDa. Panel B: SDS-PAGE ( a ) and Western blot ( b ) shows stepwise purification of hTyr. From the left: L, protein ladder; 1, total lysate of larvae expressing hTyr; 2, sample after 5 ml <t>HisTrap</t> crude column; 3, sample after Sephacryl S-300 16/60 HR SEC; 4, sample after Superdex 200 increase 10/300 GL SEC; 5, sample after Ni-NTA affinity chromatography. For the western blot anti-tyrosinase antibody (T311, 1:2000, Santa Cruz Biotechnology) was used. ( c ) shows the corresponding diphenol oxidase activity of hTyr measured after 30 min of incubation at 37°C with 3 mM L-DOPA in 50 mM sodium phosphate buffer, pH 7.4. (d) reveals the hTyr yields in protein extracts during purification, which were obtained from the SDS-PAGE gels using UN-SCAN-IT gel TM gel analysis software (Silk Scientific, Inc.).
    Ge Healthcare Histrap Crude, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ge healthcare histrap crude/product/GE Healthcare
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    ge healthcare histrap crude - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Chromatography profiles and enzymatic activities for the recombinant hTyrC tr and temperature sensitive mutant variant, R422Q A: IMAC using HisTrap FF crude 5 ml column with the linear gradient of 500 mM imidazole (gray line). B: Gel-filtration chromatography using HiPrep 16/60 Sephacryl S-100 column. The insert shows a magnification of A 280 profiles of hTyrC tr (black line) and R422Q mutant (green line). C: Gel filtration using Superdex 75 10/30 column. Top panels of A, B, and C show test tubes containing the L-DOPA colorimetric reactions for each protein fraction of hTyrC tr (black frame) and R422Q (green frame). Brown color (intensity proportional) in tube indicates diphenol oxidase activity.

    Journal: Current protocols in protein science

    Article Title: The purification of recombinant human tyrosinase from insect larvae infected with the baculovirus vector

    doi: 10.1002/cpps.37

    Figure Lengend Snippet: Chromatography profiles and enzymatic activities for the recombinant hTyrC tr and temperature sensitive mutant variant, R422Q A: IMAC using HisTrap FF crude 5 ml column with the linear gradient of 500 mM imidazole (gray line). B: Gel-filtration chromatography using HiPrep 16/60 Sephacryl S-100 column. The insert shows a magnification of A 280 profiles of hTyrC tr (black line) and R422Q mutant (green line). C: Gel filtration using Superdex 75 10/30 column. Top panels of A, B, and C show test tubes containing the L-DOPA colorimetric reactions for each protein fraction of hTyrC tr (black frame) and R422Q (green frame). Brown color (intensity proportional) in tube indicates diphenol oxidase activity.

    Article Snippet: 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.

    Techniques: Chromatography, Recombinant, Mutagenesis, Variant Assay, Filtration, Activity Assay

    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

    PstB and PhoR coelute with PhoU. (A) Genetic complementation of a Δ pstB mutation with p48 pstB and p48 pstB -His. The Δ pstB E. coli strain BW26390 was transformed with the indicated plasmids, and AP assays were performed. Cells were grown in MOPS HiP i or MOPS LoP i medium, where indicated. (B) Binding of PhoU to PstB-His. Cells were lysed, and the soluble extracts were subjected to nickel affinity purification by passage over a HisTrap column. Identical samples from the lysate and elution fraction were separated by SDS-PAGE, transferred to nitrocellulose membranes, and analyzed by immunodetection. The mixed lysate sample contained extracts from a strain expressing high levels of PstS, PstC, PstA, and PhoU with a sample expressing high levels of PstB-His. The use of the mixed lysate was necessary because the expression of PstB-His decreased the expression of the Pho regulon in the Δ pstB genetic background. (C) Plasmids expressing PhoR-His (PhoR with an amino-terminal 6×His tag) and PhoU were both transformed into a Δ phoBR Δ pstSCAB-phoU strain (SG1). Nickel affinity chromatography was used to retain the PhoR-His on the affinity column. Cell lysate and elusion fractions were analyzed by Western blotting to see that PhoU coelutes with PhoR-His.

    Journal: Journal of Bacteriology

    Article Title: The PhoU Protein from Escherichia coli Interacts with PhoR, PstB, and Metals To Form a Phosphate-Signaling Complex at the Membrane

    doi: 10.1128/JB.00029-14

    Figure Lengend Snippet: PstB and PhoR coelute with PhoU. (A) Genetic complementation of a Δ pstB mutation with p48 pstB and p48 pstB -His. The Δ pstB E. coli strain BW26390 was transformed with the indicated plasmids, and AP assays were performed. Cells were grown in MOPS HiP i or MOPS LoP i medium, where indicated. (B) Binding of PhoU to PstB-His. Cells were lysed, and the soluble extracts were subjected to nickel affinity purification by passage over a HisTrap column. Identical samples from the lysate and elution fraction were separated by SDS-PAGE, transferred to nitrocellulose membranes, and analyzed by immunodetection. The mixed lysate sample contained extracts from a strain expressing high levels of PstS, PstC, PstA, and PhoU with a sample expressing high levels of PstB-His. The use of the mixed lysate was necessary because the expression of PstB-His decreased the expression of the Pho regulon in the Δ pstB genetic background. (C) Plasmids expressing PhoR-His (PhoR with an amino-terminal 6×His tag) and PhoU were both transformed into a Δ phoBR Δ pstSCAB-phoU strain (SG1). Nickel affinity chromatography was used to retain the PhoR-His on the affinity column. Cell lysate and elusion fractions were analyzed by Western blotting to see that PhoU coelutes with PhoR-His.

    Article Snippet: The remaining cleared crude lysate was loaded onto a fresh 1-ml HisTrap FF nickel column (GE Healthcare).

    Techniques: Mutagenesis, Transformation Assay, Binding Assay, Affinity Purification, SDS Page, Immunodetection, Expressing, Affinity Chromatography, Affinity Column, Western Blot

    Recombinant hTyr purified from T . ni . larval biomass in presence of hydrogenated Triton X-100. Panel A : hTyr eluted from Superdex 200 increase 10/300 GL column after Ni-NTA gravity purification. The absorption was measured at 280 nm (black solid line) and 260 nm (black dashed line). The following Bio-Rad SEC standards are shown at the top of the panel: Thyroglobulin (670 kDa), γ-globulin (158 kDa), and ovalbumin (44 kDa). The insert shows SDS-PAGE ( a ), Western blot ( b ), and diphenol oxidase activity ( c ) for the corresponding fractions containing hTyr. Arrows display the protein ladder marker at 70 kDa. Panel B: SDS-PAGE ( a ) and Western blot ( b ) shows stepwise purification of hTyr. From the left: L, protein ladder; 1, total lysate of larvae expressing hTyr; 2, sample after 5 ml HisTrap crude column; 3, sample after Sephacryl S-300 16/60 HR SEC; 4, sample after Superdex 200 increase 10/300 GL SEC; 5, sample after Ni-NTA affinity chromatography. For the western blot anti-tyrosinase antibody (T311, 1:2000, Santa Cruz Biotechnology) was used. ( c ) shows the corresponding diphenol oxidase activity of hTyr measured after 30 min of incubation at 37°C with 3 mM L-DOPA in 50 mM sodium phosphate buffer, pH 7.4. (d) reveals the hTyr yields in protein extracts during purification, which were obtained from the SDS-PAGE gels using UN-SCAN-IT gel TM gel analysis software (Silk Scientific, Inc.).

    Journal: PLoS ONE

    Article Title: Membrane-associated human tyrosinase is an enzymatically active monomeric glycoprotein

    doi: 10.1371/journal.pone.0198247

    Figure Lengend Snippet: Recombinant hTyr purified from T . ni . larval biomass in presence of hydrogenated Triton X-100. Panel A : hTyr eluted from Superdex 200 increase 10/300 GL column after Ni-NTA gravity purification. The absorption was measured at 280 nm (black solid line) and 260 nm (black dashed line). The following Bio-Rad SEC standards are shown at the top of the panel: Thyroglobulin (670 kDa), γ-globulin (158 kDa), and ovalbumin (44 kDa). The insert shows SDS-PAGE ( a ), Western blot ( b ), and diphenol oxidase activity ( c ) for the corresponding fractions containing hTyr. Arrows display the protein ladder marker at 70 kDa. Panel B: SDS-PAGE ( a ) and Western blot ( b ) shows stepwise purification of hTyr. From the left: L, protein ladder; 1, total lysate of larvae expressing hTyr; 2, sample after 5 ml HisTrap crude column; 3, sample after Sephacryl S-300 16/60 HR SEC; 4, sample after Superdex 200 increase 10/300 GL SEC; 5, sample after Ni-NTA affinity chromatography. For the western blot anti-tyrosinase antibody (T311, 1:2000, Santa Cruz Biotechnology) was used. ( c ) shows the corresponding diphenol oxidase activity of hTyr measured after 30 min of incubation at 37°C with 3 mM L-DOPA in 50 mM sodium phosphate buffer, pH 7.4. (d) reveals the hTyr yields in protein extracts during purification, which were obtained from the SDS-PAGE gels using UN-SCAN-IT gel TM gel analysis software (Silk Scientific, Inc.).

    Article Snippet: Panel A : IMAC using a GE Healthcare HisTrap crude 5 mL column.

    Techniques: Recombinant, Purification, Size-exclusion Chromatography, SDS Page, Western Blot, Activity Assay, Marker, Expressing, Affinity Chromatography, Incubation, Software