Navigation Links
Rapid Method Development With the BioLogic DuoFlow Chromatography System for the Purification of His-Tagged Proteins

Fang-Fang Wu, Xuemei He, and David R Nau, Bio-Rad Laboratories, Inc., Hercules, CA 94547 USA


Introduction
Immobilized metal affinity chromatography (IMAC) is a powerful technique that can be used for the efficient purification of recombinant histidine-tagged (His-tagged) proteins from a variety of expression systems.

The synthesis of IMAC resins begins with the derivatization of an appropriate wide-pore base resin with metal chelating groups, such as iminodiacetic acid (IDA) or nitrilotriacetic acid (NTA). Specific transition metal ions usually Cu2+, Ni2+, Co2+, Ca2+, or Zn2+ are then immobilized to form a charged support. IMAC resins exhibit high affinity and metal-dependent selectivity for His-tagged proteins, as well as naturally occurring proteins that are rich in histidine, cysteine, aspartic acid, or glutamic acid residues.

To obtain highly purified proteins from an IMAC system, a series of method development experiments is typically required. These experiments involve optimization of the mobile phase pH, buffer system, flow rate, concentration of the competitive eluting agents, and a number of other potential parameters that may also be examined, depending on purity and recovery requirements. Usually, these would require a series of tedious and repetitive operations by the researcher, including the loading and recharging of the column with fresh metal ions after every run in order to ensure reproducible results.

Bio-Rad has developed an extremely rapid and efficient method using the BioLogic DuoFlow high-resolution chromatography system to purify a 75 kD His-tagged protein
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
2. A Simple and Rapid Technique to Purify High-Quality Midiprep DNA
3. CastAway Precast Gels for Rapid Automated DNA Sequence Analysis
4. Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
5. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
6. Simple and Rapid Isolation of Plant RNA Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit
7. Rapid PCR* Using Active Tube Temperature Control On Thermos PCR Machines
8. Molecular Biology:Easy Dephosphorylation and Rapid DNA Ligation
9. Rapid detection of genomic duplications and deletions using the Agilent 2100 bioanalyzer
10. Rapid hit confirmation and early hERG liability testing using the IonWorks HT system
11. Rapidly Prepare Probes and Markers for Small RNA Analysis
Post Your Comments:
(Date:7/10/2014)... 10, 2014 Research and Markets ( ... "International Photonic Integrated Circuit (Monolithic Integration, Hybrid ... report to their offering. ... of photonic integration traces its roots in the ... of photonic integration went unexplored and unfulfilled for ...
(Date:7/10/2014)... to the International Atomic Energy Agency (IAEA) the ... acquiring sufficient quantities of plutonium or highly enriched ... device. The IAEA also notes that most cases ... which can be challenging to detect with most ... appearing this week in the Journal of ...
(Date:7/10/2014)... D.C., June 17, 2014 Using microscopic polymer ... specific gases, researchers at MIT,s Quantum Photonics Laboratory ... levels in the parts-per-billion range. Optical sensors are ... their high signal-to-noise ratio, compact, lightweight nature, and ... gas sensors had been developed before, the MIT ...
(Date:7/10/2014)... basis of psychiatric disorders has been extremely challenging because ... but are insufficient to cause disease. Now investigators reporting ... journal Cell Stem Cell describe a strategy ... interact with other risk factors or environmental exposures to ... pinpoints a genetic variant that may predispose individuals to ...
Breaking Biology Technology:International Photonic Integrated Circuit (Monolithic Integration, Hybrid Integration, Module Integration) Market - Forecasts to 2019 2Sensitive detection method may help impede illicit nuclear trafficking 2Swell new sensors 2New strategy could uncover genes at the root of psychiatric illnesses 2
... , ROCKAWAY, N.J., Dec. 14 ... Approach , published by the Infusion Nurses ... extravasation.(1) Anthracyclines are a group of chemotherapy medications including ... the treatment of various types of cancer since the early ...
... , WALTHAM, Mass., Dec. ... that it has signed a new, three-year master services ... and services that serve patients in approximately 100 countries. ... for translation of the company,s online content, which includes ...
... WALTHAM, Mass., Dec. 14 Decision Resources, one of the world,s ... that, owing to the launches of several new agents, the atrial ... in 2008 to $6.1 billion in 2018 in the United States, ... new Pharmacor report entitled Atrial Fibrillation finds that a ...
Cached Biology Technology:Infusion Nurses Society References Totect(R) (Dexrazoxane For Injection) as the Only Antidote for the Treatment of Anthracycline Extravasation 2Infusion Nurses Society References Totect(R) (Dexrazoxane For Injection) as the Only Antidote for the Treatment of Anthracycline Extravasation 3Infusion Nurses Society References Totect(R) (Dexrazoxane For Injection) as the Only Antidote for the Treatment of Anthracycline Extravasation 4Infusion Nurses Society References Totect(R) (Dexrazoxane For Injection) as the Only Antidote for the Treatment of Anthracycline Extravasation 5Multi-Billion Dollar Biotech Firm Chooses Lionbridge as Exclusive Translation Provider with Three-Year Master Services Agreement 2Multi-Billion Dollar Biotech Firm Chooses Lionbridge as Exclusive Translation Provider with Three-Year Master Services Agreement 3The Atrial Fibrillation Drug Market Will Increase More Than Sevenfold, from $790 Million in 2008 to $6.1 Billion in 2018 2
(Date:7/11/2014)... Social animals often develop relationships with other group members ... In wild chacma baboons the strategy for grooming activities ... are just published in the scientific journal Biology ... of baboons is not practiced without ulterior motives. To ... for the individual, while grooming another individual can provide ...
(Date:7/11/2014)... 11, 2014 The increased risk of kidney injury ... resuscitation fluids reflects the mass of HES molecules, according ... official journal of the International Anesthesia Research Society (IARS). ... harmful effect of HES on cultured human renal proximal ... Christian Wunder and colleagues of University Hospital Wrzburg, Austria. ...
(Date:7/11/2014)... cannabinoid receptor type 1 can inhibit voltage-gated ... reduce neurotransmitter release. However, some scholars demonstrated ... extracellular Ca2+ influx and increase neurotransmitter release. ... Hospital Affiliated to Tongji Medical College, Huazhong ... whole cell voltage-clamp and calcium imaging in ...
Breaking Biology News(10 mins):Baboons groom early in the day to get benefits later 2In lab studies, hydroxyethyl starch has direct harmful effects on kidney cells 2
... More than 20 percent of atheist scientists are spiritual, ... public marries spirituality and religion, the study found that spirituality ... with scientific discovery for "spiritual atheist" scientists. ... Sociology of Religion . Through in-depth interviews ...
... , A new study from Cambridge University has for ... a higher than average number of autistic traits. ... today in the Journal of Autism and Developmental ... of biological girls with gender incongruence that persists into ...
... Amsterdam, 5 May 2011 Elsevier, the ... products and services, announced today the launch ... The journal aims at reporting the most recent ... intelligent computation based on the principles of swarm ...
Cached Biology News:More than 20 percent of atheist scientists are spiritual 2Female-to-male transsexual people have more autistic traits 2Elsevier announces launch of Swarm and Evolutionary Computation 2
Compound(s) tested through a broad panel of 38 ligand binding and enzyme assays targeting the Central Nervous System. Used to determine selecitivty and potential CNS-Side Effect liabilities of compo...
... tested through 63 ligand ... spanning a broad therapeutic ... includes many non human ... non-selective assays. Used ...
... Receptor (PR) Competitor Assay Kits are ... progesterone receptor binding compounds using fluorescence ... a fusion of glutathione transferase to ... progesterone receptor [PR-LBD(GST)] and a proprietary, ...
... is critical to the maintenance of ... proteins as important and diverse as ... shock and stress response, antigen presentation, ... ion channels, cell cycle regulation, transcription, ...
Biology Products: