Navigation Links
High-Level Protein Expression, One-Column Purification, and FLAG,,,Epitope Tagging in E. coli

tag for purification of proteins from E. coli extracts1 because of its high affinity for immobilized calmodulin (Kd=10-9). CBP fusion proteins are bound to and eluted from calmodulin affinity matrices at neutral pH using gentle buffer conditions. A relatively low concentration of Ca2+ ions is required for binding (0.2 to 2.0 mM CaCl2), and proteins are efficiently eluted upon removal of the calcium with 2 mM EGTA. The CBP tag is small, (4 kDa) and therefore less likely than some of the larger tags [e.g., the 26-kDa glutathione-S-transferase (GST) and 40-kDa maltose-binding protein (MBP) tags] to affect the biological function of the protein of interest. CBP is efficiently translated in E. coli, and CBP fusion proteins are typically expressed to high levels. In addition, the CBP tag is efficiently phosphorylated by protein kinase A and thus, purified CBP fusion proteins can be readily labeled with isotope and used to probe protein-protein interactions.2

Despite the advantages of using affinity tags for protein purification, there are many proteins for which fusion of nonnatural amino acids at the N- or C-termini adversely affect protein function.3 Most fusion vectors include short recognition sequences for site-specific proteases that allow removal of the fusion tag following purification. Many vectors employ target sequences for proteases that cleave internal to the recognition site and thus, leave at least one extraneous amino acid fused to the protein of interest. The use of the protease enterokinase (EK) has the advantage that it cleaves at the C-terminus of its recognition site, thus when positioned upstream of the protein of interest, N-terminal purification tags can be completely removed without leaving any extraneous amino acid residues on the protein of interest. However, most vectors that employ the use of EK for removal o
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. High-Level Expression of Peanut Allergens Affected by Rare Codon Usage
2. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
3. A New C-Terminal GST Vector for Protein Production in S. pombe
4. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
5. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
6. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
7. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
8. Optimized Imaging of Protein Gels Stained with Coomassie Brilliant Blue Dye
9. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
10. Yeast Protein Production System Features High Yields and One-Step Purification
11. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
Post Your Comments:
(Date:7/10/2014)... (PRWEB) July 10, 2014 Product ... in very low abundance and are often “lost ... and time-consuming. , Join presenters Dr. Rowel Tobias, ... Dr. John Anders, Head of Quality at Nanotherapeutics, ... approach that can speed detection and quantitation while ...
(Date:7/10/2014)... Robert Harman, DVM, Founder and CEO of Vet-Stem, Inc., the ... relaunch of his highly informative blog, now named Stem Cells ... are Stem Cells ?” Dr. Harman’s purpose for blogging ... basics of stem cell therapy so that pet owners can ... when considering regenerative medicine. , A veterinarian by trade, Dr. ...
(Date:7/10/2014)... On July 9 Prime Minister ... Russia,s first national "Industry" award ... is developing a unique project called MabNext . ... number of innovative drugs based on monoclonal antibodies for ... place at the International Exhibition "Innoprom 2014" in Ekaterinburg. ...
(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 ...
Breaking Biology Technology:Novel Ms Techniques Speed Detection of Biopharmaceutical Product and Process Impurities, New Webinar Hosted by Xtalks 2Robert Harman, DVM Talks About What Stem Cells are in His Latest Blog Series for Vet-Stem, Inc. 2Robert Harman, DVM Talks About What Stem Cells are in His Latest Blog Series for Vet-Stem, Inc. 3Dmitry Medvedev Presented BIOCAD the First National "Industry" Award 2Sensitive detection method may help impede illicit nuclear trafficking 2
... Panacea Pharmaceuticals,Inc. announced today that BC Detect(SM), ... now available from Panacea Laboratories. BC,Detect(SM) is ... the identification,of recurrence among women successfully treated ... Panacea Pharmaceuticals, is certified under the,Clinical Laboratory ...
... of anti-infectives, to ... on pexiganan development, WELLESLEY HILLS, Mass., Oct. 17 ... of rights to pexiganan, Michael Zasloff, M.D. has joined the,company,s ... the African clawed frog while at NIH (National Institutes of,Health) ...
... October 24, at 1:30 p.m. EDT, ALAMEDA, ... ), a biopharmaceutical company developing innovative therapies,for the ... call to discuss third quarter results on Wednesday, ... PDT). Kenneth Chahine, Chief Executive,Officer, and Michael Coffee, ...
Cached Biology Technology:Panacea Pharmaceuticals Announces the Availability of BC Detect(SM), a Serum- Based Diagnostic Test for Breast Cancer 2Panacea Pharmaceuticals Announces the Availability of BC Detect(SM), a Serum- Based Diagnostic Test for Breast Cancer 3Panacea Pharmaceuticals Announces the Availability of BC Detect(SM), a Serum- Based Diagnostic Test for Breast Cancer 4MacroChem Announces Michael Zasloff, M.D., Ph.D. Joins Company's Scientific Advisory Board 2MacroChem Announces Michael Zasloff, M.D., Ph.D. Joins Company's Scientific Advisory Board 3MacroChem Announces Michael Zasloff, M.D., Ph.D. Joins Company's Scientific Advisory Board 4MacroChem Announces Michael Zasloff, M.D., Ph.D. Joins Company's Scientific Advisory Board 5Avigen, Inc. Announces Third Quarter 2007 Earnings Conference Call 2
(Date:7/11/2014)... July 3, 2014, Shenzhen, China Researchers from Salk ... for the first time evaluated the safety and ... and successfully developed a new method, TALEN-HDAdV, which ... pluripotent stem cell (hiPSC). This study published online ... important theoretical foundation for stem cell-based gene therapy. ...
(Date:7/11/2014)... An international team of researchers, including scientists from the ... just a reported a major step in understanding photosynthesis, ... now maintains the oxygen in its atmosphere and which ... on earth. , The researchers report the first ... reaction, namely the step in which a specific protein ...
(Date:7/11/2014)... Researchers have pioneered a revolutionary new way to ... Virtual Finger, allows scientists to move through digital ... using the flat surface of their computer screens. ... orders of magnitude more efficient, saving time, money ... areas of experimental biology. The software and its ...
Breaking Biology News(10 mins):A new genome editing method brings the possibility of gene therapies closer to reality 2A first direct glimpse of photosynthesis in action 2Virtual finger enables scientists to navigate and analyze complex 3D images 2
... 13, 2012) The blood-brain barrier is essential for maintaining ... and isolating the brain,s specific hormonal and neurotransmitter activity from ... addition to nerve cells, the brain contains glia cells that ... blood-brain boundary is made by glia joined into an envelope ...
... DC - On January 18, 2012, Dr. Peter ... National Homeland Security Research Center in the Office ... Protection Agency, will lead a session titled "National ... national resilience to bioterrorism threats involves maximizing advanced ...
... previously un-named species of horse fly whose appearance is dominated ... honour of American pop diva, Beyonc a member of ... single, Bootylicious. , According to the Australian National Insect Collection ... ( Plinthina ) beyonceae , CSIRO,s Bryan Lessard, ...
Cached Biology News:Brain glia cells increase their DNA content to preserve vital blood-brain barrier 2Fly named in honor of Beyonce 2
HybriWell seals securely to a microscope slide surface in seconds to enclose single or multiple specimens in a small reagent volume and eliminate evaporation....
Comp Dim: chamber diam. depth 20 2.8 mm chamber maximum volume 720 μL...
... the Easy-Titer Human IgM Assay Kit to ... and easy-to-perform assay for Human IgM that ... kit incorporates a simple mix-and-read assay that ... from 15-500 ng/ml in approximately 30 minutes. ...
AC input: 230 V...
Biology Products: