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


Seamless insertion and one-step purification from a more versatile vector

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

Katherine Felts Denise Wyborski John Bauer Peter Vaillancourt
Stratagene

Stratagene has improved the Affinity LIC cloning kit with the new pCAL-n-FLAG vector. This new E. coli cloning and expression vector is designed for consistent, high-level production and one-step purification of expressed proteins. Similar to its predecessor pCAL-n-EK, pCAL-n-FLAG has the same features with the addition of the FLAG epitope ll ll as an N-terminal fusion with the protein of interest. The FLAG epitope is useful to those scientists desiring a small antibody recognition site on their protein of interest. This vector will be of particular interest to scientists who wish to use calmodulin affinity-purified proteins to study protein-protein interactions by pull-down immunoprecipitation experiments using eukaryotic cell extracts, or who wish to probe lambda cDNA expression libraries for interacting proteins. In both cases, the presence of endogenous calmodulin-binding proteins, which are ubiquitous in all eukaryotic cell types, may contribute to background. The calmodulin-binding peptide (CBP) purification tag can be removed from the recombinant protein by thrombin digestion. If desired, the FLAG tag can be removed by digestion with enterokinase, leaving protein of native amino acid sequence.

Purification of recombinant proteins has been greatly simplified in recent years due to the availability of expression vectors that allow fusion of the protein coding sequence of interest to short peptide sequences, or larger proteins, enabling the affinity purification of the fusion protein from crude preparations. The 26-amino acid CBP is an attractive fusion
'"/>

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:7/28/2014)... of building materials using light, developed by researchers at ... that are often considered the realm of science fiction, ... Although cloaked starships won,t be a reality for quite ... constructing materials with building blocks a few billionths of ... way that light flies through them, and works on ...
(Date:7/28/2014)... According to a new market research report published ... Joint) Market (By Technology - Stem Cell Therapy, ... Graft Substitutes, Osteoarticular Diseases, Allogeneic Products, Autogenic Products ... Growth, Trends and Forecast, 2013 - 2019" the ... valued at USD 2.6 billion in 2012 and ...
(Date:7/28/2014)... , July 28, 2014 ... report  "Microbial Identification Market by Products (Consumables, Instruments, Services), ... Beverages), End User (Detection, Characterization) - Global ... Microbial Identification Market is estimated at $896.5 ... reach $1,194.1 Million by 2019, growing at ...
(Date:7/28/2014)... 28, 2014 Appistry, Inc. , ... bring the power of genomics to next-generation medicine, announced ... Pipeline Challenge . , Two Appistry staff will ... National Center for Genome Resources, and Mr. Neil Miller, ... of Kansas City. Rounding out the panel are Appistry’s ...
Breaking Biology Technology:Building 'invisible' materials with light 2Regenerative Medicine Market: Global Industry Analysis, Size, Share, Growth, Trends and Forecast 2013 - 2019 2Regenerative Medicine Market: Global Industry Analysis, Size, Share, Growth, Trends and Forecast 2013 - 2019 3Regenerative Medicine Market: Global Industry Analysis, Size, Share, Growth, Trends and Forecast 2013 - 2019 4Microbial Identification Market worth $1,194.1 Million by 2019 2Microbial Identification Market worth $1,194.1 Million by 2019 3Appistry Names Industry Experts from the National Center for Genome Resources and Mercy Children's, Kansas City, to Judge Pipeline Challenge Competition 2
... Professor James Dumesic, graduate students Steven T. Evans, Tobias ... Kim. Source: UW-Madison. MADISON - Carbon monoxide, or ... the efficient operation of fuel cells. But now, chemical ... not only cleared that barrier - they also have ...
... competitors can copy the newest product enhancement, advertising ... process seemingly overnight. Indeed, it seems that no ... realization has led many corporate leaders to conclude ... is to create organizations that can respond fast ...
... General Electric Company , announced Monday it has developed ... increase efficiency for physicians. , ,The new visual stethoscope, ... larger-scale systems but in a portable and wireless design ... possible for patients to receive diagnostic exams anywhere, including ...
Cached Biology Technology:UW engineers clear bottleneck in hydrogen production 2Going beyond me, too customer experiences 2Going beyond me, too customer experiences 3Going beyond me, too customer experiences 4GE Healthcare announces new, high-tech 'stethoscope' 2
(Date:7/28/2014)... Children,s Research Hospital study found that 73 percent of ... risk of developing metabolic syndrome and related health problems ... appear in the current issue of the journal ... adult survivors of childhood cancer in the study had ... like high blood pressure, abdominal obesity, elevated triglyceride and ...
(Date:7/28/2014)... -- Research and Markets has announced the ... 2014-2018" report to their offering. ... analyze biological data. These systems capture biological traits such ... and hand measurements from the subject and compare the ... purpose of a biometrics system is to identify and ...
(Date:7/28/2014)... Researchers using statistical tools to map social connections ... traditional observational techniques, shedding light on prairie dog ... bubonic plague and guide future conservation efforts. The ... State University and the National Evolutionary Synthesis Center ... are subject to bubonic plague," says Dr. Jennifer ...
Breaking Biology News(10 mins):Unhealthy habits more than double risk of metabolic syndrome in childhood cancer survivors 2Unhealthy habits more than double risk of metabolic syndrome in childhood cancer survivors 3Biometrics Market in Latin America 2014-2018 2Social network research may boost prairie dog conservation efforts 2
... -- Chronic water quality problems caused by agricultural ... the burning of fossil fuels leads to oxygen depletion, ... pollution is prevalent in many coastal marine and estuarine ... North America and although many efforts have been taken ...
... has revealed the mechanism that binds skin cells tightly ... for painful and debilitating skin diseases and also lethal ... of Life Sciences, has found that the glue molecules ... making a very tough, resilient structure. Further investigation on ...
... can be an opportunity for researchers to foster respectful ... This shift is proposed in a policy forum appearing ... of the American Academy for the Advancement of Science. ... Practices and Participant Preferences: The Growing Gulf" recommend new ...
Cached Biology News:Could oysters be used to clean up Chesapeake Bay? 2Cell binding discovery brings hope to those with skin and heart problems 2Gulf grows between research practice and participant preferences in genetic studies 2Gulf grows between research practice and participant preferences in genetic studies 3Gulf grows between research practice and participant preferences in genetic studies 4
... Adhesion Molecule The L1CAM ... involved in three distinct conditions: 1) HSAS ... MASA (mental retardation, aphasia, shuffling gait, adductus ... Immunogen: Homogenous suspension of ...
Immunogen: C terminus [VQLKRSKNDSKPYC] of cardiac elav-type RNA-binding protein (ETR-3) Storage: -20 C, Avoid Freeze/Thaw Cycles...
... PP2A1, purified Formulation: 50mM ... 0.02% Brij-35, 0.2mM PMSF, 1mM ... glycerol. Properites: PP2A1 (Protein Phosphatase ... DEAE-Sephadex, ammonium sulfate precipitation, QSepharose,amino-hexyl ...
Although each Beacon 2000 System includes one preinstalled sample chamber (size specified at time of order), optional chambers are available and easily retrofit by the user....
Biology Products: