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TaKaRa talon superflow metal affinity resin
Enrichment of chloroplast-derived cholera toxin-B subunit (CTB) malarial antigens. (a) CTB FC apical membrane antigen-1 (AMA1) protein was extracted from transformed leaves and the crude extract was subjected to <t>Talon</t> <t>Superflow</t> Metal Affinity Resin and
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1) Product Images from "Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery"

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery

Journal:

doi: 10.1111/j.1467-7652.2009.00479.x

Enrichment of chloroplast-derived cholera toxin-B subunit (CTB) malarial antigens. (a) CTB FC apical membrane antigen-1 (AMA1) protein was extracted from transformed leaves and the crude extract was subjected to Talon Superflow Metal Affinity Resin and
Figure Legend Snippet: Enrichment of chloroplast-derived cholera toxin-B subunit (CTB) malarial antigens. (a) CTB FC apical membrane antigen-1 (AMA1) protein was extracted from transformed leaves and the crude extract was subjected to Talon Superflow Metal Affinity Resin and

Techniques Used: Derivative Assay, CtB Assay, Flow Cytometry, Transformation Assay

2) Product Images from "Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery"

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery

Journal:

doi: 10.1111/j.1467-7652.2009.00479.x

Enrichment of chloroplast-derived cholera toxin-B subunit (CTB) malarial antigens. (a) CTB FC apical membrane antigen-1 (AMA1) protein was extracted from transformed leaves and the crude extract was subjected to Talon Superflow Metal Affinity Resin and
Figure Legend Snippet: Enrichment of chloroplast-derived cholera toxin-B subunit (CTB) malarial antigens. (a) CTB FC apical membrane antigen-1 (AMA1) protein was extracted from transformed leaves and the crude extract was subjected to Talon Superflow Metal Affinity Resin and

Techniques Used: Derivative Assay, CtB Assay, Flow Cytometry, Transformation Assay

3) Product Images from "Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic"

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic

Journal:

doi: 10.1074/jbc.M110.127605

Purification of the recombinant Tay1 protein. Tay1p-6HN was purified by TALON Superflow metal affinity chromatography (heparin input), loaded onto a heparin column, and eluted by a linear 0–1.5 m NaCl gradient, and the proteins were analyzed by 10% SDS-PAGE. Shown are peak fractions eluted from the heparin column. Fractions 10–16 were pooled, dialyzed, and used for subsequent experiments.
Figure Legend Snippet: Purification of the recombinant Tay1 protein. Tay1p-6HN was purified by TALON Superflow metal affinity chromatography (heparin input), loaded onto a heparin column, and eluted by a linear 0–1.5 m NaCl gradient, and the proteins were analyzed by 10% SDS-PAGE. Shown are peak fractions eluted from the heparin column. Fractions 10–16 were pooled, dialyzed, and used for subsequent experiments.

Techniques Used: Purification, Recombinant, Affinity Chromatography, SDS Page

4) Product Images from "Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytica"

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytica

Journal:

doi: 10.1074/jbc.M110.127605

Purification of the recombinant Tay1 protein. Tay1p-6HN was purified by TALON Superflow metal affinity chromatography (heparin input), loaded onto a heparin column, and eluted by a linear 0–1.5 m NaCl gradient, and the proteins were analyzed by 10% SDS-PAGE. Shown are peak fractions eluted from the heparin column. Fractions 10–16 were pooled, dialyzed, and used for subsequent experiments.
Figure Legend Snippet: Purification of the recombinant Tay1 protein. Tay1p-6HN was purified by TALON Superflow metal affinity chromatography (heparin input), loaded onto a heparin column, and eluted by a linear 0–1.5 m NaCl gradient, and the proteins were analyzed by 10% SDS-PAGE. Shown are peak fractions eluted from the heparin column. Fractions 10–16 were pooled, dialyzed, and used for subsequent experiments.

Techniques Used: Purification, Recombinant, Affinity Chromatography, SDS Page

Related Articles

Clone Assay:

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Briefly, DNAs encoding the polypeptides were constructed from synthetic oligonucleotides using PCR and cloned into the expression vector pET32b+ (Merck KGaA, Darmstadt, Germany). .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: We synthesized N-terminal His-tagged YGFP derivatives, GFPuv and EGFP by standard PCR techniques and cloned them into the pEGFP vector at the Hind III and EcoR I sites, as described above. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: DNA fragments encoding the polypeptides were cloned into the expression vector pET32b+ (Novagen, USA). .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ].

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Paragraph title: Cloning and expression of EqCXCL16 in Escherichia coli . ... The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

Centrifugation:

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic
Article Snippet: The insoluble material was pelleted by 30 min of centrifugation at 12,000 rpm at 4 °C (F21–8 × 50y in Sorvall RC 6+). .. The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A.

Article Title: Generation of a More Immunogenic Measles Vaccine by Increasing Its Hemagglutinin Expression
Article Snippet: Supernatants and cells were collected at 24-h intervals and lysed by one freeze-thaw cycle, and supernatants were clarified by centrifugation at 3,000 × g for 5 min in an SX4750 rotor. .. Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction.

Article Title: Creation of a type IIS restriction endonuclease with a long recognition sequence
Article Snippet: Thirty microliters of 50% slurry of TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA), pre-equilibrated with 50 mM sodium phosphate, pH 7.0, 0.5 M NaCl and 50 mM MgCl2 , were added to each, and the mixtures were rocked for an hour at 4°C. .. Thirty microliters of 50% slurry of TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA), pre-equilibrated with 50 mM sodium phosphate, pH 7.0, 0.5 M NaCl and 50 mM MgCl2 , were added to each, and the mixtures were rocked for an hour at 4°C.

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: After freezing at −80 °C and thawing, 5 µg/ml DNase I, 10 µg/ml RNase A and 10 mM MgCl2 were added to the bacterial lysate. .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole. .. After extensive washes at 10 mM then 30 mM imidazole in the same buffer, the H-HadD protein was finally eluted using 150 mM imidazole (Supplementary Fig. ).

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: Cell suspensions were lysed in a microfluidizer processor M-110EH (Microfluidics). .. Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump. .. Bound protein was washed with about 10 column volumes in the corresponding lysis buffer followed by a wash with about five column volumes in buffer containing 0.3 M NaCl, 20 mM HEPES pH 7.5, 5 mM imidazole, 0.05% sodium azide.

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: The bacterial pellet was re-suspended in lysis buffer (20 mM Tris-HCl buffer, pH 8.0, 150 mM NaCl, 1 mM phenylmethylsulphonyl fluoride (PMSF), 5% (v/v) glycerol and 1 mg mL-1 lysozyme, and then lysed on ice using a Sonopuls ultrasonic homogenizer with a 6-mm diameter probe (50% duty cycle; amplitude setting, 30%; total time, 30 s, 6–8 times). .. Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow. .. The column was washed with 8 column volumes (cv) of equilibration buffer (20 mM Tris-HCl buffer, pH 8.0 containing 150 mM NaCl), followed by 7 cv of the equilibration buffer containing 10 mM imidazole.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Following centrifugation at 6,000 × g for 15 min, the cell pellet was resuspended in buffer A (50 mM sodium phosphate, 6 M guanidine-HCl, 300 mM NaCl, pH 7.0) and subjected to several short cycles of sonication to reduce viscosity. .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

Amplification:

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: The MSMEG_0948 gene from M . smegmatis mc²155 was amplified by PCR and the product was ligated into pET-15b vector (Novagen) between Nde I and Bam HI restriction sites; the absence of mutation was verified by sequencing. .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Filtration:

Article Title: Bacillus subtilis Intramembrane Protease RasP Activity in Escherichia coli and In Vitro
Article Snippet: The supernatant was mixed with 1 ml Talon Superflow metal affinity resin (Clontech) that had been equilibrated with buffer (PBS containing 1% DM, 5 mM 2-mercaptoethanol, and 10% glycerol). .. The supernatant was mixed with 1 ml Talon Superflow metal affinity resin (Clontech) that had been equilibrated with buffer (PBS containing 1% DM, 5 mM 2-mercaptoethanol, and 10% glycerol).

Mass Spectrometry:

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA). .. The recombinant APHC1/3 polypeptides were purified on a reverse-phase column Jupiter C4 (Phenomenex, Torrance, CA, USA) 250 × 10 mm.

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol. .. In order to isolate the properly folded recombinant peptide, the reaction mixture was subjected to two separation steps: (1) RP-HPLC using a Jupiter C5 (250 × 10 mm, Phenomenex, Torrance, CA, USA) column and a linear gradient from 0% to 60% MeCN over 60 min (flow of 10 mL/min); and (2) RP-HPLC using a Vydac C18 (250 × 4.6 mm, Grace, Williamsburg, MI, USA) column and a linear gradient from 0% to 40% MeCN over 40 min (flow of 1 mL/min).

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ]. .. The recombinant polypeptides were purified on a reverse-phase column Jupiter C5 (Phenomenex, USA) 250×10 mm.

Article Title: Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity
Article Snippet: The reaction was terminated by adding 5X SDS sample loading buffer (10% SDS, 25% β-mercaptoethanol, 50% glycerol, 0.125 M Tris-HCl, pH6.8) and heating at 95C for 5 min. After SilverSNAP staining, a gel slice containing SR7:9 was excised and processed for mass spec analysis; pY1123 was the most abundant site identified, along with Y963, Y1000, Y1017, Y1165 and Y1211. .. His6 -bKal7 , expressed in Hi5 cells was purified using Talon Superflow Metal Affinity Resin (#635506; Clontech Laboratories) equilibrated with 50 mM Na phosphate, 300 mM NaCl, 0.05% TX-100, pH 8.0, eluted with a gradient to 1.0 M imidazole in the same buffer titrated to pH 7.0 and dialyzed into 20 mM Na TES, 0.3 M NaCl, pH 8.0.

Synthesized:

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: We synthesized N-terminal His-tagged YGFP derivatives, GFPuv and EGFP by standard PCR techniques and cloned them into the pEGFP vector at the Hind III and EcoR I sites, as described above. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Construct:

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Briefly, DNAs encoding the polypeptides were constructed from synthetic oligonucleotides using PCR and cloned into the expression vector pET32b+ (Merck KGaA, Darmstadt, Germany). .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: For expression of the N-terminal His6 -tagged HadD protein, E . coli RosettaTM (DE3) strain (Novagen) was transformed by pET-15b:: MSMEG_0948 construct and grown at 37 °C under shaking (200 rpm) in Terrific broth medium (Euromedex) supplemented with carbenicillin (50 µg/ml) and chloramphenicol (30 µg/ml). .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: One-liter cultures were induced with 0.5 mM isopropyl-β-d-thiogalactopyranoside (IPTG) at an optical density of 0.8 at 37°C for 4 h. Bacterial cells were suspended in either 1 M NaCl, 25 mM Tris–HCl pH 7.5, 1 mM EDTA, 0.05% sodium azide for the constructs containing the maltose binding protein (see above) or 1 M NaCl, 20 mM HEPES pH 7.5, 5 mM Imidazole, 0.05% sodium azide for the RRM. .. Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL). .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

Enzyme-linked Immunosorbent Assay:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: To address whether CTB fusion protein generated in transplastomic tobacco and lettuce retained biological activities and could bind to the GM1 receptor, we performed GM1 -binding ELISA assay. .. Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

Incubation:

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic
Article Snippet: Triton X-100 was added to a final concentration of 0.1%, and the suspension was sonicated one more time for 30 s and incubated for additional 15 min on ice. .. The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A.

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: To detect CTB-AMA1 and CTB-MSP1-fused proteins, blots were incubated with 1 : 3000 rabbit anti-CTB primary polyclonal antibody (Sigma, St Louis, MO, USA) followed by 1 : 5000 HRP-conjugated donkey anti-rabbit secondary antibody (Southernbiotech, Birmingham, AL, USA). .. The supernatant was collected and then subjected to TALON superflow Metal Affinity Resin (Clontech) to enrich the chloroplast-derived CTB-malarial proteins according to manufacturer’s instructions.

Article Title: Generation of a More Immunogenic Measles Vaccine by Increasing Its Hemagglutinin Expression
Article Snippet: Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction. .. Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction.

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: After overnight incubation, cells were resuspended in PBS containing cOmplete™ Mini EDTA-free protease-inhibitor cocktail (Roche, Basel, Switzerland) and sonicated for 3 min (30 sec x 6 pulses) on ice with Q500 (Qsonica, LLC. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C. .. After incubation, the resin was washed 5 times with PBS and then washed in PBS containing 200 mM imidazole to elute His-tagged proteins.

Activity Assay:

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow. .. Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow.

Cell Culture:

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Escherichia coli BL21(DE3) cells expressing TRX fusions of APHC1 or APHC3 were cultured at 25 °C for 12–14 h, harvested, resuspended in start buffer for metal-affinity chromatography (300 mM NaCl, 50 mM Tris-HCl, pH 7.5), ultrasonicated, and centrifuged to remove all insoluble particles. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: Escherichia coli BL21 (DE3) cells expressing thioredoxin fusions of polypeptides were cultured overnight at 25°C, harvested, ultrasonicated in a buffer for metal-affinity chromatography, and centrifuged to remove all insoluble particles. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ].

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: The cell culture was cooled to 20°C, before isopropyl thio-β -D-galactoside (IPTG) was added to a final concentration of 0.4 mM for GlcNAcase expression. .. Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow.

Article Title: Recombinant phosphatidylserine-binding nanobodies for targeting of extracellular vesicles to tumor cells: a plug-and-play approach †Electronic supplementary information (ESI) available. See DOI: 10.1039/c7nr06966a
Article Snippet: Sepharose CL-4B and cholesterol were obtained from Sigma-Aldrich (Steinheim, Germany) and TALON Superflow Metal Affinity Resin was from Clontech Laboratories, Inc. (Saint-Germain-en-Laye, France). .. Sepharose CL-4B and cholesterol were obtained from Sigma-Aldrich (Steinheim, Germany) and TALON Superflow Metal Affinity Resin was from Clontech Laboratories, Inc. (Saint-Germain-en-Laye, France).

Expressing:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: Paragraph title: Expression, quantification and enrichment of vaccine antigens ... Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

Article Title: A viral scaffolding protein triggers portal ring oligomerization and incorporation during procapsid assembly
Article Snippet: To purify PMs, plasmid pET21b containing full-length gene 1 (portal protein) ( ) was transformed into Escherichia coli strain BL21 (DE3) cells for expression. .. The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag.

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Escherichia coli BL21(DE3) cells expressing TRX fusions of APHC1 or APHC3 were cultured at 25 °C for 12–14 h, harvested, resuspended in start buffer for metal-affinity chromatography (300 mM NaCl, 50 mM Tris-HCl, pH 7.5), ultrasonicated, and centrifuged to remove all insoluble particles. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: Creation of a type IIS restriction endonuclease with a long recognition sequence
Article Snippet: Paragraph title: Screen for high expression level of I-SceI mutant proteins in E. coli ... Thirty microliters of 50% slurry of TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA), pre-equilibrated with 50 mM sodium phosphate, pH 7.0, 0.5 M NaCl and 50 mM MgCl2 , were added to each, and the mixtures were rocked for an hour at 4°C.

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: Expression was induced at a culture density of ~0.6–0.8 (OD600 ) by adding isopropyl-1-thio-β-d -galactopyranoside up to 0.2 mM. .. The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol.

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: Cultures were stopped at an OD600 of 0.8 and left at 13 °C for 30 min. Gene expression was induced by addition of 1 mM IPTG, and bacteria were grown further for 24 h at 13 °C and 200 rpm. .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Article Title: A Novel Antibiotic Mechanism of l-Cyclopropylalanine Blocking the Biosynthetic Pathway of Essential Amino Acid l-Leucine
Article Snippet: Paragraph title: 3.5. Protein Expression and Purification ... The bacterial cells were harvested and protein purification was carried out by Immobilized Metal ion Affinity Column (IMAC) method using Talon SuperFlow Metal Affinity Resin (Clontech, Mountain View, CA, USA) as recommended by the manufacturer.

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: Paragraph title: Bacterial protein expression and purification ... Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump.

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: Paragraph title: Expression and purification of YGFP derivatives and other GFP proteins in E . coli ... The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: Escherichia coli BL21 (DE3) cells expressing thioredoxin fusions of polypeptides were cultured overnight at 25°C, harvested, ultrasonicated in a buffer for metal-affinity chromatography, and centrifuged to remove all insoluble particles. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ].

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: Paragraph title: Protein expression and purification ... Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Paragraph title: Cloning and expression of EqCXCL16 in Escherichia coli . ... The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

BIA-KA:

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C. .. To determine protein concentration, purified proteins dissolved in PBS were diluted 2-fold with 10% SDS and then incubated at 95°C for 10 min.

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow. .. Fractions with GlcNAcase activity were pooled and subjected to several rounds of centrifugation in Vivaspin-20 ultrafiltration membrane concentrators (10 kDa molecular-weight cut-off, Vivascience AG, Hannover, Germany) for complete removal of imidazole.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ). .. The purity and integrity of recombinant EqCXCL16 (aa 17 to 247) were evaluated by subjecting it to electrophoresis on a 4 to 20% gradient gel-SDS followed by staining with Coomassie brilliant blue solution (Bio-Rad Laboratories, Hercules, CA).

Acrylamide Gel Assay:

Article Title: Generation of a More Immunogenic Measles Vaccine by Increasing Its Hemagglutinin Expression
Article Snippet: Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction. .. Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction.

Western Blot:

Article Title: Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity
Article Snippet: His6 -bKal7 , expressed in Hi5 cells was purified using Talon Superflow Metal Affinity Resin (#635506; Clontech Laboratories) equilibrated with 50 mM Na phosphate, 300 mM NaCl, 0.05% TX-100, pH 8.0, eluted with a gradient to 1.0 M imidazole in the same buffer titrated to pH 7.0 and dialyzed into 20 mM Na TES, 0.3 M NaCl, pH 8.0. .. Based on the rosette screen and the biologically significant role played by Kalirin in TrkB signaling , a subset of 12 GST-SH2 and -PTB domains were tested for their ability to bind to bKal7 phosphorylated in vitro by recombinant TrkB kinase.

Transformation Assay:

Article Title: A viral scaffolding protein triggers portal ring oligomerization and incorporation during procapsid assembly
Article Snippet: To purify PMs, plasmid pET21b containing full-length gene 1 (portal protein) ( ) was transformed into Escherichia coli strain BL21 (DE3) cells for expression. .. The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag.

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: The expression vector was transformed into competent cells, which were then cultivated at 37 °C in LB medium containing ampicillin at a concentration of 100 µg/mL. .. The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol.

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: For expression of the N-terminal His6 -tagged HadD protein, E . coli RosettaTM (DE3) strain (Novagen) was transformed by pET-15b:: MSMEG_0948 construct and grown at 37 °C under shaking (200 rpm) in Terrific broth medium (Euromedex) supplemented with carbenicillin (50 µg/ml) and chloramphenicol (30 µg/ml). .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: The constructs were transformed in Rosetta™ 2 (DE3) pLysS cell (Novagen). .. Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump.

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: Briefly, the transformed cells were grown at 37°C in Terrific Broth (TB) containing 100 μg mL-1 ampicillin and 25 μg mL-1 kanamycin until the cell density reached an OD600 of 0.6. .. Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: In order to express recombinant protein, the plasmid was transformed into E. coli BL21(DE3) cells (EMD Millipore Novagen, Temecula, CA). .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

Flow Cytometry:

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol. .. HCl was added to a final concentration of 0.2 M, and the CNBr molar ratio to peptide was adjusted to 600:1.

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: Cell suspensions were lysed in a microfluidizer processor M-110EH (Microfluidics). .. Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump. .. Bound protein was washed with about 10 column volumes in the corresponding lysis buffer followed by a wash with about five column volumes in buffer containing 0.3 M NaCl, 20 mM HEPES pH 7.5, 5 mM imidazole, 0.05% sodium azide.

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: The bacterial pellet was re-suspended in lysis buffer (20 mM Tris-HCl buffer, pH 8.0, 150 mM NaCl, 1 mM phenylmethylsulphonyl fluoride (PMSF), 5% (v/v) glycerol and 1 mg mL-1 lysozyme, and then lysed on ice using a Sonopuls ultrasonic homogenizer with a 6-mm diameter probe (50% duty cycle; amplitude setting, 30%; total time, 30 s, 6–8 times). .. Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow. .. The column was washed with 8 column volumes (cv) of equilibration buffer (20 mM Tris-HCl buffer, pH 8.0 containing 150 mM NaCl), followed by 7 cv of the equilibration buffer containing 10 mM imidazole.

Article Title: Rapid and Scalable Plant-based Production of a Cholera Toxin B Subunit Variant to Aid in Mass Vaccination against Cholera Outbreaks
Article Snippet: Talon Superflow Metal Affinity Resin (Clontech), packed in an XK-26 column (GE Healthcare) to a 50 ml bed volume, was equilibrated with 10 column volumes (CV) of buffer A (20 mM Tris-Cl, pH 8.0, 500 mM NaCl). .. Talon Superflow Metal Affinity Resin (Clontech), packed in an XK-26 column (GE Healthcare) to a 50 ml bed volume, was equilibrated with 10 column volumes (CV) of buffer A (20 mM Tris-Cl, pH 8.0, 500 mM NaCl).

Chromatography:

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Escherichia coli BL21(DE3) cells expressing TRX fusions of APHC1 or APHC3 were cultured at 25 °C for 12–14 h, harvested, resuspended in start buffer for metal-affinity chromatography (300 mM NaCl, 50 mM Tris-HCl, pH 7.5), ultrasonicated, and centrifuged to remove all insoluble particles. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: Escherichia coli BL21 (DE3) cells expressing thioredoxin fusions of polypeptides were cultured overnight at 25°C, harvested, ultrasonicated in a buffer for metal-affinity chromatography, and centrifuged to remove all insoluble particles. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ].

Article Title: Rapid and Scalable Plant-based Production of a Cholera Toxin B Subunit Variant to Aid in Mass Vaccination against Cholera Outbreaks
Article Snippet: Chromatography was performed using an AKTA Purifier (GE Healthcare). .. Talon Superflow Metal Affinity Resin (Clontech), packed in an XK-26 column (GE Healthcare) to a 50 ml bed volume, was equilibrated with 10 column volumes (CV) of buffer A (20 mM Tris-Cl, pH 8.0, 500 mM NaCl).

Protease Inhibitor:

Article Title: Creation of a type IIS restriction endonuclease with a long recognition sequence
Article Snippet: The cell pellets were recovered by centrifugation (10 min at 3220g , 4°C), stored frozen at –20°C, then thawed on ice and lysed in 350 μl of 50 mM sodium phosphate, pH 7.0, 0.5 M NaCl, 50 mM MgCl2 , 0.5 mg/ml lysozyme, 0.05 mg/ml DNaseI, and 1× EDTA-free COMPLETE protease inhibitor cocktail (Roche, Indianapolis, IN). .. Thirty microliters of 50% slurry of TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA), pre-equilibrated with 50 mM sodium phosphate, pH 7.0, 0.5 M NaCl and 50 mM MgCl2 , were added to each, and the mixtures were rocked for an hour at 4°C.

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: After overnight incubation, cells were resuspended in PBS containing cOmplete™ Mini EDTA-free protease-inhibitor cocktail (Roche, Basel, Switzerland) and sonicated for 3 min (30 sec x 6 pulses) on ice with Q500 (Qsonica, LLC. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Buffer Exchange:

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C. .. After incubation, the resin was washed 5 times with PBS and then washed in PBS containing 200 mM imidazole to elute His-tagged proteins.

Infection:

Article Title: Generation of a More Immunogenic Measles Vaccine by Increasing Its Hemagglutinin Expression
Article Snippet: To isolate 6×His-tagged soluble H protein from the supernatants of MVvac2-Hsol-infected cells, 100-mm-diameter dishes were seeded with 1 × 106 Vero/hSLAM cells and infected with MVvac2 or MVvac2-Hsol at an MOI of 0.1. .. Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction.

SDS Page:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: The protein separated by SDS-PAGE gel was transferred to nitrocellulose membrane by electroblotting and the membrane was blocked overnight with 3% non-fat dry milk. .. The supernatant was collected and then subjected to TALON superflow Metal Affinity Resin (Clontech) to enrich the chloroplast-derived CTB-malarial proteins according to manufacturer’s instructions.

Article Title: A viral scaffolding protein triggers portal ring oligomerization and incorporation during procapsid assembly
Article Snippet: The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag. .. The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag.

Article Title: Generation of a More Immunogenic Measles Vaccine by Increasing Its Hemagglutinin Expression
Article Snippet: Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction. .. Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction.

Article Title: Creation of a type IIS restriction endonuclease with a long recognition sequence
Article Snippet: Thirty microliters of 50% slurry of TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA), pre-equilibrated with 50 mM sodium phosphate, pH 7.0, 0.5 M NaCl and 50 mM MgCl2 , were added to each, and the mixtures were rocked for an hour at 4°C. .. Thirty microliters of 50% slurry of TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA), pre-equilibrated with 50 mM sodium phosphate, pH 7.0, 0.5 M NaCl and 50 mM MgCl2 , were added to each, and the mixtures were rocked for an hour at 4°C.

Article Title: A Novel Antibiotic Mechanism of l-Cyclopropylalanine Blocking the Biosynthetic Pathway of Essential Amino Acid l-Leucine
Article Snippet: The bacterial cells were harvested and protein purification was carried out by Immobilized Metal ion Affinity Column (IMAC) method using Talon SuperFlow Metal Affinity Resin (Clontech, Mountain View, CA, USA) as recommended by the manufacturer. .. The bacterial cells were harvested and protein purification was carried out by Immobilized Metal ion Affinity Column (IMAC) method using Talon SuperFlow Metal Affinity Resin (Clontech, Mountain View, CA, USA) as recommended by the manufacturer.

Article Title: Rapid and Scalable Plant-based Production of a Cholera Toxin B Subunit Variant to Aid in Mass Vaccination against Cholera Outbreaks
Article Snippet: Talon Superflow Metal Affinity Resin (Clontech), packed in an XK-26 column (GE Healthcare) to a 50 ml bed volume, was equilibrated with 10 column volumes (CV) of buffer A (20 mM Tris-Cl, pH 8.0, 500 mM NaCl). .. The CTB was eluted with a step gradient using 100% buffer B (Buffer A+150 mM imidazole) and collected by monitoring absorbance at 280 nm.

Sedimentation:

Article Title: A viral scaffolding protein triggers portal ring oligomerization and incorporation during procapsid assembly
Article Snippet: After induction, cells were harvested by sedimentation in a Sorvall SLC-6000 rotor at 5368g for 15 min, resuspended in binding buffer [20 mM Hepes (pH 7.5), 500 mM NaCl, and 10 mM imidazole], and frozen at −20°C. .. The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag.

Generated:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: To address whether CTB fusion protein generated in transplastomic tobacco and lettuce retained biological activities and could bind to the GM1 receptor, we performed GM1 -binding ELISA assay. .. Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

Protein Concentration:

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ). .. The purity and integrity of recombinant EqCXCL16 (aa 17 to 247) were evaluated by subjecting it to electrophoresis on a 4 to 20% gradient gel-SDS followed by staining with Coomassie brilliant blue solution (Bio-Rad Laboratories, Hercules, CA).

Polymerase Chain Reaction:

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Briefly, DNAs encoding the polypeptides were constructed from synthetic oligonucleotides using PCR and cloned into the expression vector pET32b+ (Merck KGaA, Darmstadt, Germany). .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: The MSMEG_0948 gene from M . smegmatis mc²155 was amplified by PCR and the product was ligated into pET-15b vector (Novagen) between Nde I and Bam HI restriction sites; the absence of mutation was verified by sequencing. .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: We synthesized N-terminal His-tagged YGFP derivatives, GFPuv and EGFP by standard PCR techniques and cloned them into the pEGFP vector at the Hind III and EcoR I sites, as described above. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Sonication:

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic
Article Snippet: Triton X-100 was added to a final concentration of 0.1%, and the suspension was sonicated one more time for 30 s and incubated for additional 15 min on ice. .. The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A.

Article Title: A viral scaffolding protein triggers portal ring oligomerization and incorporation during procapsid assembly
Article Snippet: The cells were thawed on ice, followed by the addition of phenylmethylsulfonyl fluoride (1 mM), deoxyribonuclease (50 μg/ml), ribonuclease (50 μg/ml), MgCl2 (2 mM), and CaCl2 (0.5 mM), and lysed by sonication at 35 A, 15-s pulse, and 30-s pause, with a total processing time of 3 min using a Misonix Sonicator 4000 with a standard tip. .. The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag.

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: After overnight incubation, cells were resuspended in PBS containing cOmplete™ Mini EDTA-free protease-inhibitor cocktail (Roche, Basel, Switzerland) and sonicated for 3 min (30 sec x 6 pulses) on ice with Q500 (Qsonica, LLC. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Following centrifugation at 6,000 × g for 15 min, the cell pellet was resuspended in buffer A (50 mM sodium phosphate, 6 M guanidine-HCl, 300 mM NaCl, pH 7.0) and subjected to several short cycles of sonication to reduce viscosity. .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

Affinity Purification:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: Cholera toxin was shown to bind to immobilized Ni2+ ion mediated by the B subunit and is a useful affinity purification method for CTB and complexes composed of CTB ( ). .. Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

Binding Assay:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: These results suggest that the tobacco and lettuce chloroplast-derived CTB-AMA1 and CTB-MSP1 assemble properly to form pentameric structures, which are essential for GM1 -ganglioside receptor binding. .. Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

Article Title: A viral scaffolding protein triggers portal ring oligomerization and incorporation during procapsid assembly
Article Snippet: After induction, cells were harvested by sedimentation in a Sorvall SLC-6000 rotor at 5368g for 15 min, resuspended in binding buffer [20 mM Hepes (pH 7.5), 500 mM NaCl, and 10 mM imidazole], and frozen at −20°C. .. The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag.

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: One-liter cultures were induced with 0.5 mM isopropyl-β-d-thiogalactopyranoside (IPTG) at an optical density of 0.8 at 37°C for 4 h. Bacterial cells were suspended in either 1 M NaCl, 25 mM Tris–HCl pH 7.5, 1 mM EDTA, 0.05% sodium azide for the constructs containing the maltose binding protein (see above) or 1 M NaCl, 20 mM HEPES pH 7.5, 5 mM Imidazole, 0.05% sodium azide for the RRM. .. Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump.

Article Title: Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity
Article Snippet: Paragraph title: Far-Western analysis of GST-SH2 binding to TrkB phosphorylated bKal7 ... His6 -bKal7 , expressed in Hi5 cells was purified using Talon Superflow Metal Affinity Resin (#635506; Clontech Laboratories) equilibrated with 50 mM Na phosphate, 300 mM NaCl, 0.05% TX-100, pH 8.0, eluted with a gradient to 1.0 M imidazole in the same buffer titrated to pH 7.0 and dialyzed into 20 mM Na TES, 0.3 M NaCl, pH 8.0.

Molecular Weight:

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow. .. Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow.

In Vivo:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: GM1 -ganglioside has been shown to be the receptor for CTB protein in vivo ( ) and a pentameric structure is required for binding to GM1 receptor ( ). .. Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

Mutagenesis:

Article Title: Creation of a type IIS restriction endonuclease with a long recognition sequence
Article Snippet: Paragraph title: Screen for high expression level of I-SceI mutant proteins in E. coli ... Thirty microliters of 50% slurry of TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA), pre-equilibrated with 50 mM sodium phosphate, pH 7.0, 0.5 M NaCl and 50 mM MgCl2 , were added to each, and the mixtures were rocked for an hour at 4°C.

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: The MSMEG_0948 gene from M . smegmatis mc²155 was amplified by PCR and the product was ligated into pET-15b vector (Novagen) between Nde I and Bam HI restriction sites; the absence of mutation was verified by sequencing. .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Isolation:

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Recombinant plasmid p15-16A (aa 17 to 247) was isolated from ampicillin-resistant clones using a ZR plasmid miniprep kit (Zymo Research, Irvine, CA). .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

Size-exclusion Chromatography:

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: After overnight incubation, cells were resuspended in PBS containing cOmplete™ Mini EDTA-free protease-inhibitor cocktail (Roche, Basel, Switzerland) and sonicated for 3 min (30 sec x 6 pulses) on ice with Q500 (Qsonica, LLC. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Purification:

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic
Article Snippet: Paragraph title: Purification of Recombinant Tay1 Protein ... The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A.

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic
Article Snippet: The column was then calibrated using (50 μl of 1 mg/ml solution each) blue dextran (void volume), apoferritin (440 kDa), aldolase (147 kDa), and bovine serum albumin (67 kDa). .. After calibration, 50 μl of 0.2 mg/ml Tay1p purified by TALON Superflow Metal Affinity Resin was loaded on the column, and 0.25-ml fractions were collected starting from the void volume. .. Fractions adsorbing UV light were analyzed by 10% SDS-PAGE ( ).

Article Title: A viral scaffolding protein triggers portal ring oligomerization and incorporation during procapsid assembly
Article Snippet: Cell debris was removed by centr ifugation in a Sorvall F18-12 × 50 rotor at 38,725g for 15 min. .. The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag. .. Unbound protein was eluted with wash buffer [20 mM Hepes (pH 7.5), 500 mM NaCl, and 20 mM imidazole].

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Escherichia coli BL21(DE3) cells expressing TRX fusions of APHC1 or APHC3 were cultured at 25 °C for 12–14 h, harvested, resuspended in start buffer for metal-affinity chromatography (300 mM NaCl, 50 mM Tris-HCl, pH 7.5), ultrasonicated, and centrifuged to remove all insoluble particles. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA). .. HCl up to 0.5 M and CNBr with a molar ratio to protein of 600:1 were added to the fusion protein solution in the metal-affinity chromatography elution buffer [ ].

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: After cultivation, the cells were harvested by centrifuging for 5 min at 6000× g , re-suspended in buffer for metal affinity chromatography (400 mM NaCl, 20 mM Tris-HCl, pH 7.5), ultrasonicated, and centrifuged for 15 min at 9000× g to remove all insoluble particles. .. The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol. .. The fusion protein was diluted to 1 mg/mL and cleaved overnight with CNBr in the dark at room temperature, as previously described [ ].

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: Paragraph title: Protein production and purification ... After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Article Title: A Novel Antibiotic Mechanism of l-Cyclopropylalanine Blocking the Biosynthetic Pathway of Essential Amino Acid l-Leucine
Article Snippet: Paragraph title: 3.5. Protein Expression and Purification ... The bacterial cells were harvested and protein purification was carried out by Immobilized Metal ion Affinity Column (IMAC) method using Talon SuperFlow Metal Affinity Resin (Clontech, Mountain View, CA, USA) as recommended by the manufacturer.

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: Paragraph title: Bacterial protein expression and purification ... Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump.

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: Paragraph title: Expression and purification of YGFP derivatives and other GFP proteins in E . coli ... The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: Escherichia coli BL21 (DE3) cells expressing thioredoxin fusions of polypeptides were cultured overnight at 25°C, harvested, ultrasonicated in a buffer for metal-affinity chromatography, and centrifuged to remove all insoluble particles. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ]. .. The recombinant polypeptides were purified on a reverse-phase column Jupiter C5 (Phenomenex, USA) 250×10 mm.

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: Paragraph title: Protein expression and purification ... Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow.

Article Title: Rapid and Scalable Plant-based Production of a Cholera Toxin B Subunit Variant to Aid in Mass Vaccination against Cholera Outbreaks
Article Snippet: Paragraph title: Purification of CTB ... Talon Superflow Metal Affinity Resin (Clontech), packed in an XK-26 column (GE Healthcare) to a 50 ml bed volume, was equilibrated with 10 column volumes (CV) of buffer A (20 mM Tris-Cl, pH 8.0, 500 mM NaCl).

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Following centrifugation at 6,000 × g for 15 min, the cell pellet was resuspended in buffer A (50 mM sodium phosphate, 6 M guanidine-HCl, 300 mM NaCl, pH 7.0) and subjected to several short cycles of sonication to reduce viscosity. .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ). .. The columns were equilibrated and washed with buffer A, while proteins were eluted using buffer B (45 mM sodium phosphate, 5.4 M guanidine-HCl, 270 mM NaCl, 150 mM imidazole, pH 7.0).

Article Title: Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity
Article Snippet: The reaction was terminated by adding 5X SDS sample loading buffer (10% SDS, 25% β-mercaptoethanol, 50% glycerol, 0.125 M Tris-HCl, pH6.8) and heating at 95C for 5 min. After SilverSNAP staining, a gel slice containing SR7:9 was excised and processed for mass spec analysis; pY1123 was the most abundant site identified, along with Y963, Y1000, Y1017, Y1165 and Y1211. .. His6 -bKal7 , expressed in Hi5 cells was purified using Talon Superflow Metal Affinity Resin (#635506; Clontech Laboratories) equilibrated with 50 mM Na phosphate, 300 mM NaCl, 0.05% TX-100, pH 8.0, eluted with a gradient to 1.0 M imidazole in the same buffer titrated to pH 7.0 and dialyzed into 20 mM Na TES, 0.3 M NaCl, pH 8.0. .. Purified bKal7 was phosphorylated as described above but using 100 nM TrkB and 200 nM bKal7 with 4 mM ATP; TrkB was not added to the buffer control.

Protein Purification:

Article Title: A Novel Antibiotic Mechanism of l-Cyclopropylalanine Blocking the Biosynthetic Pathway of Essential Amino Acid l-Leucine
Article Snippet: Isopropyl-1-thio-β-d -galactoside (IPTG) was added to the culture to the final concentration of 0.5 mM. .. The bacterial cells were harvested and protein purification was carried out by Immobilized Metal ion Affinity Column (IMAC) method using Talon SuperFlow Metal Affinity Resin (Clontech, Mountain View, CA, USA) as recommended by the manufacturer. .. The protein was dialyzed (to remove immidazole) and concentrated using 3 kDa Amicon Ultra centrifugal filter (Millipore, Burlington, MA, USA), and kept in 1× equilibrium buffer (50 mM Sodium phosphate, 300 mM NaCl, pH 7.0) and 50% glycerol.

Article Title: Bacillus subtilis Intramembrane Protease RasP Activity in Escherichia coli and In Vitro
Article Snippet: Paragraph title: Protein purification. ... The supernatant was mixed with 1 ml Talon Superflow metal affinity resin (Clontech) that had been equilibrated with buffer (PBS containing 1% DM, 5 mM 2-mercaptoethanol, and 10% glycerol).

Sequencing:

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol. .. In order to isolate the properly folded recombinant peptide, the reaction mixture was subjected to two separation steps: (1) RP-HPLC using a Jupiter C5 (250 × 10 mm, Phenomenex, Torrance, CA, USA) column and a linear gradient from 0% to 60% MeCN over 60 min (flow of 10 mL/min); and (2) RP-HPLC using a Vydac C18 (250 × 4.6 mm, Grace, Williamsburg, MI, USA) column and a linear gradient from 0% to 40% MeCN over 40 min (flow of 1 mL/min).

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: The MSMEG_0948 gene from M . smegmatis mc²155 was amplified by PCR and the product was ligated into pET-15b vector (Novagen) between Nde I and Bam HI restriction sites; the absence of mutation was verified by sequencing. .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Recombinant:

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic
Article Snippet: Paragraph title: Purification of Recombinant Tay1 Protein ... The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A.

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Paragraph title: 3.1. Production of Recombinant Polypeptides ... Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: Paragraph title: 5.6. Recombinant Peptide Production ... The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol.

Article Title: A Novel Antibiotic Mechanism of l-Cyclopropylalanine Blocking the Biosynthetic Pathway of Essential Amino Acid l-Leucine
Article Snippet: E. coli BL21 (λDE3) cells harboring recombinant plasmids were grown at 37 °C in LB medium supplemented with 100 μg/mL ampicillin until the culture reached mid log phase (0.3–0.4, OD600 ). .. The bacterial cells were harvested and protein purification was carried out by Immobilized Metal ion Affinity Column (IMAC) method using Talon SuperFlow Metal Affinity Resin (Clontech, Mountain View, CA, USA) as recommended by the manufacturer.

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: Paragraph title: Production of recombinant polypeptides ... Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ].

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Following centrifugation at 6,000 × g for 15 min, the cell pellet was resuspended in buffer A (50 mM sodium phosphate, 6 M guanidine-HCl, 300 mM NaCl, pH 7.0) and subjected to several short cycles of sonication to reduce viscosity. .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ). .. The columns were equilibrated and washed with buffer A, while proteins were eluted using buffer B (45 mM sodium phosphate, 5.4 M guanidine-HCl, 270 mM NaCl, 150 mM imidazole, pH 7.0).

Article Title: Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity
Article Snippet: His6 -bKal7 , expressed in Hi5 cells was purified using Talon Superflow Metal Affinity Resin (#635506; Clontech Laboratories) equilibrated with 50 mM Na phosphate, 300 mM NaCl, 0.05% TX-100, pH 8.0, eluted with a gradient to 1.0 M imidazole in the same buffer titrated to pH 7.0 and dialyzed into 20 mM Na TES, 0.3 M NaCl, pH 8.0. .. Phosphorylated bKal7 was prepared for mass spec analysis as described for SR7:9; pY1123 was the major site identified.

Affinity Chromatography:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: Cholera toxin was shown to bind to immobilized Ni2+ ion mediated by the B subunit and is a useful affinity purification method for CTB and complexes composed of CTB ( ). .. Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions. .. The large subunit of Rubisco (55 kDa) was present in the untransformed, lysate and flow through fractions under reduced and non-reduced conditions ( ).

Article Title: Generation of a More Immunogenic Measles Vaccine by Increasing Its Hemagglutinin Expression
Article Snippet: Paragraph title: Immobilized cobalt ion affinity chromatography. ... Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction.

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: After cultivation, the cells were harvested by centrifuging for 5 min at 6000× g , re-suspended in buffer for metal affinity chromatography (400 mM NaCl, 20 mM Tris-HCl, pH 7.5), ultrasonicated, and centrifuged for 15 min at 9000× g to remove all insoluble particles. .. The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Following centrifugation at 6,000 × g for 15 min, the cell pellet was resuspended in buffer A (50 mM sodium phosphate, 6 M guanidine-HCl, 300 mM NaCl, pH 7.0) and subjected to several short cycles of sonication to reduce viscosity. .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ). .. The columns were equilibrated and washed with buffer A, while proteins were eluted using buffer B (45 mM sodium phosphate, 5.4 M guanidine-HCl, 270 mM NaCl, 150 mM imidazole, pH 7.0).

Fast Protein Liquid Chromatography:

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic
Article Snippet: The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A. .. The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A.

Spectroscopy:

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump. .. Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump.

Lysis:

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: Cell suspensions were lysed in a microfluidizer processor M-110EH (Microfluidics). .. Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump. .. Bound protein was washed with about 10 column volumes in the corresponding lysis buffer followed by a wash with about five column volumes in buffer containing 0.3 M NaCl, 20 mM HEPES pH 7.5, 5 mM imidazole, 0.05% sodium azide.

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: The bacterial pellet was re-suspended in lysis buffer (20 mM Tris-HCl buffer, pH 8.0, 150 mM NaCl, 1 mM phenylmethylsulphonyl fluoride (PMSF), 5% (v/v) glycerol and 1 mg mL-1 lysozyme, and then lysed on ice using a Sonopuls ultrasonic homogenizer with a 6-mm diameter probe (50% duty cycle; amplitude setting, 30%; total time, 30 s, 6–8 times). .. Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow.

Liquid Chromatography:

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: Following centrifugation at 6,000 × g for 15 min, the cell pellet was resuspended in buffer A (50 mM sodium phosphate, 6 M guanidine-HCl, 300 mM NaCl, pH 7.0) and subjected to several short cycles of sonication to reduce viscosity. .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ). .. The columns were equilibrated and washed with buffer A, while proteins were eluted using buffer B (45 mM sodium phosphate, 5.4 M guanidine-HCl, 270 mM NaCl, 150 mM imidazole, pH 7.0).

Plasmid Preparation:

Article Title: A viral scaffolding protein triggers portal ring oligomerization and incorporation during procapsid assembly
Article Snippet: To purify PMs, plasmid pET21b containing full-length gene 1 (portal protein) ( ) was transformed into Escherichia coli strain BL21 (DE3) cells for expression. .. The supernatant was loaded onto a 15-ml Talon Superflow metal affinity resin (Clontech) for purification of the portal protein via the engineered C-terminal hexahistidine (His6 ) tag.

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: Briefly, DNAs encoding the polypeptides were constructed from synthetic oligonucleotides using PCR and cloned into the expression vector pET32b+ (Merck KGaA, Darmstadt, Germany). .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: The expression vector was transformed into competent cells, which were then cultivated at 37 °C in LB medium containing ampicillin at a concentration of 100 µg/mL. .. The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol.

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: The MSMEG_0948 gene from M . smegmatis mc²155 was amplified by PCR and the product was ligated into pET-15b vector (Novagen) between Nde I and Bam HI restriction sites; the absence of mutation was verified by sequencing. .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: We synthesized N-terminal His-tagged YGFP derivatives, GFPuv and EGFP by standard PCR techniques and cloned them into the pEGFP vector at the Hind III and EcoR I sites, as described above. .. The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C.

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: DNA fragments encoding the polypeptides were cloned into the expression vector pET32b+ (Novagen, USA). .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ].

Article Title: Recombinant phosphatidylserine-binding nanobodies for targeting of extracellular vesicles to tumor cells: a plug-and-play approach †Electronic supplementary information (ESI) available. See DOI: 10.1039/c7nr06966a
Article Snippet: MicroBCA Protein Assay Kit, AlexaFluor 488 NHS ester, CellTracker Green CMFDA dye, CellTracker Deep Red dye, CellTracker Red CMTPX dye and the pcDNA3.1 vector with Neomycin resistance were purchased from Thermo Fisher Scientific (Waltham, USA). .. Sepharose CL-4B and cholesterol were obtained from Sigma-Aldrich (Steinheim, Germany) and TALON Superflow Metal Affinity Resin was from Clontech Laboratories, Inc. (Saint-Germain-en-Laye, France).

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: In order to express recombinant protein, the plasmid was transformed into E. coli BL21(DE3) cells (EMD Millipore Novagen, Temecula, CA). .. The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

Electrophoresis:

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ). .. Eluted recombinant protein was dialyzed against PBS using Slide-A-Lyser dialysis cassettes (Thermo Scientific, Rockford, IL).

Functional Assay:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: As illustrated in , transplastomic CTB-AMA1 and CTB-MSP1 protein are fully functional and binds to GM1 . .. Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

Positron Emission Tomography:

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: For expression of the N-terminal His6 -tagged HadD protein, E . coli RosettaTM (DE3) strain (Novagen) was transformed by pET-15b:: MSMEG_0948 construct and grown at 37 °C under shaking (200 rpm) in Terrific broth medium (Euromedex) supplemented with carbenicillin (50 µg/ml) and chloramphenicol (30 µg/ml). .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

In Vitro:

Article Title: Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity
Article Snippet: His6 -bKal7 , expressed in Hi5 cells was purified using Talon Superflow Metal Affinity Resin (#635506; Clontech Laboratories) equilibrated with 50 mM Na phosphate, 300 mM NaCl, 0.05% TX-100, pH 8.0, eluted with a gradient to 1.0 M imidazole in the same buffer titrated to pH 7.0 and dialyzed into 20 mM Na TES, 0.3 M NaCl, pH 8.0. .. Phosphorylated bKal7 was prepared for mass spec analysis as described for SR7:9; pY1123 was the major site identified.

Nuclear Magnetic Resonance:

Article Title: The structural basis of CstF-77 modulation of cleavage and polyadenylation through stimulation of CstF-64 activity
Article Snippet: Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump. .. Lysates were clarified by centrifugation at 16 000 × g for 20 min at 4°C and loaded on home-packed columns either with an Amylose High Flow resin (New England Biolabs) or TALON Superflow Metal Affinity resin (TaKaRa) pre-equilibrated with the corresponding lysis buffers using peristaltic pump.

CtB Assay:

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: Cholera toxin was shown to bind to immobilized Ni2+ ion mediated by the B subunit and is a useful affinity purification method for CTB and complexes composed of CTB ( ). .. Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Article Snippet: The samples were placed on ice and homogenized for 5 min with an OMNI International (GLH-2596) probe and centrifuged at 700 g for 15 min at 4 °C. .. The supernatant was collected and then subjected to TALON superflow Metal Affinity Resin (Clontech) to enrich the chloroplast-derived CTB-malarial proteins according to manufacturer’s instructions. .. The eluted fraction was collected and subjected to the Bradford Protein assay (BioRad) and to the RC-DC Protein Assay (Bio-Rad).

Article Title: Rapid and Scalable Plant-based Production of a Cholera Toxin B Subunit Variant to Aid in Mass Vaccination against Cholera Outbreaks
Article Snippet: Paragraph title: Purification of CTB ... Talon Superflow Metal Affinity Resin (Clontech), packed in an XK-26 column (GE Healthcare) to a 50 ml bed volume, was equilibrated with 10 column volumes (CV) of buffer A (20 mM Tris-Cl, pH 8.0, 500 mM NaCl).

Produced:

Article Title: Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia
Article Snippet: APHC1 and APHC3 were produced as described previously [ , , ]. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, Mountain View, CA, USA).

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: The recombinant Ueq 12-1 was produced as a fusion protein with a thioredoxin domain using an Escherichia coli SHuffle® T7 Express strain (New England Biolabs, Ipswich, MA, USA). .. The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol.

Article Title: TRPV1 activation power can switch an action mode for its polypeptide ligands
Article Snippet: Polypeptides APHC1-3 were produced as previously described [ ]. .. Fusion proteins were purified using a TALON Superflow Metal Affinity Resin (Clontech, USA) and subjected to CNBr cleavage, as described [ ].

Concentration Assay:

Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic
Article Snippet: Triton X-100 was added to a final concentration of 0.1%, and the suspension was sonicated one more time for 30 s and incubated for additional 15 min on ice. .. The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A.

Article Title: New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties
Article Snippet: The expression vector was transformed into competent cells, which were then cultivated at 37 °C in LB medium containing ampicillin at a concentration of 100 µg/mL. .. The fusion protein was purified using the TALON Superflow metal affinity resin (Clontech, Mountain View, CA, USA) according to the manufacturer’s protocol.

Article Title: HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness
Article Snippet: After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole. .. After centrifugation at 27,000 g for 20 min at 4 °C, the supernatant was loaded onto TALON® superflow metal affinity resin (Clontech) equilibrated with 50 mM HEPES buffer pH 7.5, 300 mM NaCl and 10 mM imidazole.

Article Title: A Novel Antibiotic Mechanism of l-Cyclopropylalanine Blocking the Biosynthetic Pathway of Essential Amino Acid l-Leucine
Article Snippet: Isopropyl-1-thio-β-d -galactoside (IPTG) was added to the culture to the final concentration of 0.5 mM. .. The bacterial cells were harvested and protein purification was carried out by Immobilized Metal ion Affinity Column (IMAC) method using Talon SuperFlow Metal Affinity Resin (Clontech, Mountain View, CA, USA) as recommended by the manufacturer.

Article Title: Molecular evolution of versatile derivatives from a GFP-like protein in the marine copepod Chiridius poppei
Article Snippet: The soluble protein fraction was mixed with TALON Superflow Metal Affinity Resin (Takara Bio Inc) and incubated overnight at 4°C. .. After incubation, the resin was washed 5 times with PBS and then washed in PBS containing 200 mM imidazole to elute His-tagged proteins.

Article Title: Probing the Catalytic Mechanism of Vibrio harveyi GH20 β-N-Acetylglucosaminidase by Chemical Rescue
Article Snippet: The cell culture was cooled to 20°C, before isopropyl thio-β -D-galactoside (IPTG) was added to a final concentration of 0.4 mM for GlcNAcase expression. .. Unbroken cells and cell debris were removed by centrifugation at 12,000 ×g for 1 h. The supernatant containing Vh GlcNAcase was immediately applied to a polypyrene column packed with 5 mL of TALON® Superflow™ metal affinity resin (Clontech Laboratories, Inc., USA) operated at 4°C with gravity-dependent flow.

Staining:

Article Title: Generation of a More Immunogenic Measles Vaccine by Increasing Its Hemagglutinin Expression
Article Snippet: Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction. .. Talon Superflow metal affinity resin (Clontech Laboratories, Mountain View, CA) equilibrated in interaction buffer (50 mM NaH2 PO4 [pH 8], 200 mM NaCl, 1% Triton X-100) was added to the clarified supernatants for 2 h of batch interaction.

Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
Article Snippet: The His-tagged recombinant EqCXCL16 protein (aa 17 to 247) was purified from the supernatant by affinity chromatography using Talon Superflow metal affinity resin (Clontech Laboratories Inc., Mountain View, CA) in combination with a fast performance liquid chromatography apparatus (Amersham Pharmacia Biotech Inc., Piscataway, NJ). .. Eluted recombinant protein was dialyzed against PBS using Slide-A-Lyser dialysis cassettes (Thermo Scientific, Rockford, IL).

Article Title: Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity
Article Snippet: The reaction was terminated by adding 5X SDS sample loading buffer (10% SDS, 25% β-mercaptoethanol, 50% glycerol, 0.125 M Tris-HCl, pH6.8) and heating at 95C for 5 min. After SilverSNAP staining, a gel slice containing SR7:9 was excised and processed for mass spec analysis; pY1123 was the most abundant site identified, along with Y963, Y1000, Y1017, Y1165 and Y1211. .. His6 -bKal7 , expressed in Hi5 cells was purified using Talon Superflow Metal Affinity Resin (#635506; Clontech Laboratories) equilibrated with 50 mM Na phosphate, 300 mM NaCl, 0.05% TX-100, pH 8.0, eluted with a gradient to 1.0 M imidazole in the same buffer titrated to pH 7.0 and dialyzed into 20 mM Na TES, 0.3 M NaCl, pH 8.0.

Affinity Column:

Article Title: A Novel Antibiotic Mechanism of l-Cyclopropylalanine Blocking the Biosynthetic Pathway of Essential Amino Acid l-Leucine
Article Snippet: Isopropyl-1-thio-β-d -galactoside (IPTG) was added to the culture to the final concentration of 0.5 mM. .. The bacterial cells were harvested and protein purification was carried out by Immobilized Metal ion Affinity Column (IMAC) method using Talon SuperFlow Metal Affinity Resin (Clontech, Mountain View, CA, USA) as recommended by the manufacturer. .. The protein was dialyzed (to remove immidazole) and concentrated using 3 kDa Amicon Ultra centrifugal filter (Millipore, Burlington, MA, USA), and kept in 1× equilibrium buffer (50 mM Sodium phosphate, 300 mM NaCl, pH 7.0) and 50% glycerol.

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    TaKaRa talon superflow metal affinity resin
    Purification of the recombinant Tay1 protein. Tay1p-6HN was purified by <t>TALON</t> <t>Superflow</t> metal affinity chromatography (heparin input), loaded onto a heparin column, and eluted by a linear 0–1.5 m NaCl gradient, and the proteins were analyzed by 10% SDS-PAGE. Shown are peak fractions eluted from the heparin column. Fractions 10–16 were pooled, dialyzed, and used for subsequent experiments.
    Talon Superflow Metal Affinity Resin, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Purification of the recombinant Tay1 protein. Tay1p-6HN was purified by TALON Superflow metal affinity chromatography (heparin input), loaded onto a heparin column, and eluted by a linear 0–1.5 m NaCl gradient, and the proteins were analyzed by 10% SDS-PAGE. Shown are peak fractions eluted from the heparin column. Fractions 10–16 were pooled, dialyzed, and used for subsequent experiments.

    Journal:

    Article Title: Tay1 Protein, a Novel Telomere Binding Factor from Yarrowia lipolytic

    doi: 10.1074/jbc.M110.127605

    Figure Lengend Snippet: Purification of the recombinant Tay1 protein. Tay1p-6HN was purified by TALON Superflow metal affinity chromatography (heparin input), loaded onto a heparin column, and eluted by a linear 0–1.5 m NaCl gradient, and the proteins were analyzed by 10% SDS-PAGE. Shown are peak fractions eluted from the heparin column. Fractions 10–16 were pooled, dialyzed, and used for subsequent experiments.

    Article Snippet: The supernatant was mixed with a 1-ml bed volume of the TALON Superflow Metal Affinity Resin (Clontech) equilibrated with 3 × 10 volumes of buffer A.

    Techniques: Purification, Recombinant, Affinity Chromatography, SDS Page

    Enrichment of chloroplast-derived cholera toxin-B subunit (CTB) malarial antigens. (a) CTB FC apical membrane antigen-1 (AMA1) protein was extracted from transformed leaves and the crude extract was subjected to Talon Superflow Metal Affinity Resin and

    Journal:

    Article Title: Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery

    doi: 10.1111/j.1467-7652.2009.00479.x

    Figure Lengend Snippet: Enrichment of chloroplast-derived cholera toxin-B subunit (CTB) malarial antigens. (a) CTB FC apical membrane antigen-1 (AMA1) protein was extracted from transformed leaves and the crude extract was subjected to Talon Superflow Metal Affinity Resin and

    Article Snippet: Therefore, for enrichment crude plant extract was subjected to immobilized metal affinity chromatography using the TALON Superflow Metal Affinity Resin (Clontech) and run on NuPAGE Novex Bis– Tris gradient gel under reducing and non-reducing conditions.

    Techniques: Derivative Assay, CtB Assay, Flow Cytometry, Transformation Assay