histrap ff crude lysate columns  (GE Healthcare)

 
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    HisTrap FF
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    HisTrap FF
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    GE Healthcare histrap ff crude lysate columns

    https://www.bioz.com/result/histrap ff crude lysate columns/product/GE Healthcare
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    histrap ff crude lysate columns - by Bioz Stars, 2021-01
    99/100 stars

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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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  • 99
    GE Healthcare histrap ff nickel column
    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 <t>HisTrap</t> 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.
    Histrap Ff Nickel Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histrap ff nickel column/product/GE Healthcare
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    histrap ff nickel column - by Bioz Stars, 2021-01
    99/100 stars
      Buy from Supplier

    99
    GE Healthcare histrap crude ff column
    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 <t>HisTrap</t> 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.
    Histrap Crude Ff Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histrap crude ff column/product/GE Healthcare
    Average 99 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    histrap crude ff column - by Bioz Stars, 2021-01
    99/100 stars
      Buy from Supplier

    92
    GE Healthcare histrap ff crude column
    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 <t>HisTrap</t> 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.
    Histrap Ff Crude Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histrap ff crude column/product/GE Healthcare
    Average 92 stars, based on 245 article reviews
    Price from $9.99 to $1999.99
    histrap ff crude column - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

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    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