Purification of the RelB and RelE Proteins of Escherichia ,

In a control experiment (data not shown), the 33-kDa GST-RelB fusion was purified from extracts of BL21(DE3) carrying only plasmid pJT4 by affinity chromatography on glutathione columns (Pharmacia) This purified protein did not adsorb to a nickel affinity column...

Nickel Columns and Nickel Resin | Bio-Rad

Nickel beads can be used in low- and medium-pressure chromatography systems or in gravity flow and spin columns Bio-Rad's Nickel-Charged Columns, Resins, and Sensor Chips In addition to nickel column chromatography, nickel-NTA can also be used to capture histidine-tagged proteins on a surface for surface plasmon resonance (SPR) analysis...

What happens if the Ni-NTA resin on my column runs dry?

What happens if the Ni-NTA resin on my column runs dry? , With BioRad or Pharmacia-type columns you can even completely remove the resin (with ,...

A high-copy T7 Escherichia coli expression vector for the ,

A T7 E coli plasmid able to express a minimal His-tag Brazilian Journal of Medical and Biological Research (2004) 37: 1103-1109 ISSN 0100-879X A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide 1Centro de Biotecnologia, Instituto Butantan, São Paulo ....

A high-copy T7 Escherichia coli expression vector for the ,

A T7 E coli plasmid able to express a minimal His-tag Brazilian Journal of Medical and Biological Research (2004) 37: 1103-1109 ISSN 0100-879X A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide 1Centro de Biotecnologia, Instituto Butantan, São Paulo ....

Ni-NTA Purification System - Thermo Fisher Scientific

Ni-NTA Purification System with Antibody, with Anti-His(C-term)-HRP Antibody K953-01 Ni-NTA Agarose (10 mL) R901-01 Ni-NTA Agarose (25 mL) R901-15 Ni-NTA Agarose (100 mL) R901-10 System Components The Ni-NTA Purification System components are listed in the following table and include enough resin, reagents, and columns for six purifications...

SUPPLEMENTARY METHODS Trojan peptide technology

on nickel columns upon expression in Escherichia coli The sequence coding for an epitope recognized by a specific antibody, between restriction sites NheI and BamHI, was replaced by the sequence coding for TAT 16 aa peptide, derived form the 86-amino acid Tat transactivation...

p47 is a cofactor for p97-mediated membrane fusion | Nature

The full-length cDNA was modified to include a His 6 tag at the N terminus and the tagged p47 was expressed and purified on nickel columns (Fig 2a, lane 3) Recombinant His 6-p47 still bound p97 ....

Fusions and display libraries

When you are ready to purify your protein product, you take advantage of the fact that consecutive histidines can join forces to bind divalent cations such as nickel You can apply your lysate to a nickel chelation resin, where the his-tagged protein will stick, and wash away the ,...

Pharmacia, PhotoWorks and Enron - MarketWatch

Applied Digital Solutions ADSX rose 8 cents, or 25 percent, to 40 cents on heavy volume of 369 million shares, putting it in fourth place on the Nasdaq volume chart behind heavyweights such as ....

Activation of a mitogen-activated protein kinase pathway ,

His-tagged proteins were purified by using nickel columns (Amersham Pharmacia), desalted, and concentrated by using Centricon-10 (Millipore) Antibody Preparation and Immunoblot Analysis NtMEK2 antibody was raised in rabbit against purified recombinant protein cor-responding to the N-terminal 156 amino acids of NtMEK2 that...

Purification of Recombinant Arabidopsis thaliana Dehydrins ,

Re- ibrated with water and charged with 04 ml of 3 mg/ml suspended proteins were desalted on Sephadex G-25 F of the respective metal salt followed by washing with in prepacked PD-10 columns (Pharmacia) using Buff- 10 column volumes of water...

Compatible-Solute-Supported Periplasmic Expression of ,

desalting columns (Pharmacia) Recombinant immunotoxin was partially puri-fied by immobilized metal ion affinity chromatography (IMAC) with nickel-nitriloacetic acid (Ni21-NTA) chelating Sepharose (Qiagen) on a BioLogic workstation (Bio-Rad, Munich, Germany) Bound protein was eluted with 250...

Yeast Sco1, a protein essential for cytochrome c oxidase ,

3 Abstract Sco1 is a conserved, essential protein, which has been implicated in the delivery of copper to cytochrome c oxidase, the last enzyme of the electron transport chain In this study, we show for the first time that the purified C-terminal domain of yeast Sco1 binds one Cu(I) per monomer...

Activation of a mitogen-activated protein kinase pathway ,

Jan 16, 2001· BL21(DE3) cells transformed with pET-28a(+) constructs were induced with 05 mM isopropyl β-D-thiogalactoside for 3 h His-tagged proteins were purified by using nickel columns (Amersham Pharmacia), desalted, and concentrated by using Centricon-10 (Millipore)...

Fast protein liquid chromatography - Wikipedia

Fast protein liquid chromatography (FPLC), is a form of liquid chromatography that is often used to analyze or purify mixtures of proteins As in other forms of chromatography, separation is possible because the different components of a mixture have different affinities for two materials, a moving fluid (the "mobile phase") and a porous solid (the stationary phase)...

Engineered Protein Polymer-Gold Nanoparticle Hybrid ,

were purchased from Amersham Pharmacia Biotech AB (Piscataway, NJ) Columns were purchased from Bio-Rad (Hercules, CA) Site-directed mutagenesis pQE30/CE1 and pQE30/E1C were employed for production of CE1-His6 and E1C-His6 , IMAC FF column charged with nickel under denaturing conditions For the negative control....

Nickel Resin | ge healthcare | Bioz

His-FLAG-tagged XerS and XerS Y341F were purified on nickel resin (1 ml His-trap HP, GE Healthcare) followed by heparin (1 ml High-Trap HP, GE Healthcare) and gel filtration (HiLoad 16/60 Superdex 200, GE Healthcare) columns ( Supplementary Figure S1 ) Purified recombinases were concentrated with an ICON concentrator (Thermoscientific) and ....

Nickel Resin | ge healthcare | Bioz

His-FLAG-tagged XerS and XerS Y341F were purified on nickel resin (1 ml His-trap HP, GE Healthcare) followed by heparin (1 ml High-Trap HP, GE Healthcare) and gel filtration (HiLoad 16/60 Superdex 200, GE Healthcare) columns ( Supplementary Figure S1 ) Purified recombinases were concentrated with an ICON concentrator (Thermoscientific) and ....

FPLC Columns | Labx

FPLC Columns This column chromatography system automates the separation and purification of semi-preparative amounts of the compound of interest, typically a protein The system uses aqueous buffers and fairly low pressures, and is housed in a "cold box" so that chromatographic runs can be done at 4°C, thus helping to preserve biological ....

Immobilization of engineered proteins on a nickel ,

Issue in Honor of Prof Alain Krief ARKIVOC 2007 (x) 268-277 Immobilization of engineered proteins on a nickel chelating molecular wire Fabrice Balavoine,a Anne Girard-Nguyen,a Patrick Schultz,b Jacques Haiech,c and Charles Mioskowskia,* a CEA-Saclay, Service de Marquage Moléculaire et de Chimie Bioorganique, Bâtiment 547, 91191 Gif sur Yvette cedex, France...

THE JOURNAL OF BIOLOGICAL CHEMISTRY - jbcorg

(Novagen), and PKR was purified over nickel columns as previously described (34) His-tagged proteins were further purified on a gel fil-tration column, Superose 12 (Amersham Pharmacia Biotech), examined by Coomassie Blue-stained SDS-polyacrylamide gels ,...

Research & Biopharmaceutical Manufacturing - GE Healthcare ,

We offer solutions to support your work from biological research to clinical therapy, including tools for research, drug discovery, diagnostics and bioprocessing...

Escherichia coli HypA Is a Zinc Metalloprotein with a Weak ,

The hyp operon encodes accessory proteins that are required for the maturation of the [NiFe] hydrogenase enzymes and, in some organisms, for the production of urease enzymes as well HypA or a homologous protein is required for nickel insertion into the hydrogenase precursor proteins In this study, recombinant HypA from Escherichia coli was purified and characterized in vitro...

Chromatography Columns | Life Science Research | Bio-Rad

Glass Econo-Column ® Columns High-quality, low-pressure, glass chromatography columns ranging in size from 5 to 170 cm long and 05 to 50 cm in diameter Glass Econo-Column Accessories Reservoirs are available in 500 ml and 1 L capacities and fit 05, 07, 10, or 15 cm ID Econo-Column Chromatography Columns...

Protocol for Histag based Ni-NTA protein purification

Protocol for Histag based Ni-NTA protein purification , There is also on-line an old manual for GST-fusion from Pharmacia or Amersham , I reached the SDS wash and my columns started running ....

Purification of his-tagged proteins in non-denaturing ,

which would allow the his-tagging system to be used for similar protein interaction studi As previously described (9, 10, 11), a histidine-tagged recombinant protein can be purified with commercially available nickel chelating resins (Qiagen, Pharmacia), by eluting bound proteins at low pH (<50) We have modified the T7 expression...

Nur, a nickel-responsive regulator of the Fur family ,

About Cookies, including instructions on how to turn off cookies if you wish to do so By continuing to browse this site you agree to us using cookies as described in ,...

TheStressOncoproteinLEDGF/p75InteractswiththeMethyl CpG ,

at 4 C for 1 hour His-LEDGF/p75 or His-p52 were then incubatedwiththebeadsat4 Cfor1hourinHEPESbuffer The beads were then collected by centrifugation at 5,000 rpm for 30 seconds at 4 C, the supernatant was discharged, and the beads were washed 2 times with 1 mL of HEPES buffer þ 01% NP-40 followed by HEPES buffer þ 05% NP-40 þ 05 mol/L NaCl...

HiTrap Columns - Sigma-Aldrich

HiTrap Desalting columns are ready-to-use 5mL HiTrap columns, pre-packed with Sephadex® G-25 Superfine for group separa-tions, buffer exchange, and removal of low molecular weight contaminat Up to 5 columns connected in series can be operated with a ,...