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:8/21/2014)... 21, 2014  BioSpecifics Technologies Corp. (NASDAQ: ... in class collagenase-based products marketed as XIAFLEX ® ... U.S. and XIAPEX ® in the EU, ... randomized, double-blind Phase 2a study of CCH for ... panniculopathy. The results showed that all three doses ...
(Date:8/21/2014)... , August 21, 2014 ... http://www.researchandmarkets.com/research/3zh3q8/global_hormone ) has announced the addition of ...  report to their offering.       ... as hormone therapy, is a means of ... produced by the body. This type of ...
(Date:8/21/2014)... Mathematic studies at the Program for Evolutionary Dynamics ... Jeffrey Epstein VI Foundation , have shown that the ... elimination. These aggressive cells are the driver mutations that drive ... of what to look for when examining tumors, comes at ... can be extracted from biopsy, it,s very difficult to know ...
(Date:8/21/2014)... Sofia, Bulgaria (PRWEB) August 21, 2014 ... was released today by Ontotext . Now ... and semantic analysis of structured and unstructured data is ... of the cost of enterprise technology. Organizations that do ... approaches for data management can use S4 since there ...
Breaking Biology Technology:BioSpecifics Technologies Corp. Announces Positive Data from Phase 2a Study of CCH for Treatment of Cellulite 2BioSpecifics Technologies Corp. Announces Positive Data from Phase 2a Study of CCH for Treatment of Cellulite 3BioSpecifics Technologies Corp. Announces Positive Data from Phase 2a Study of CCH for Treatment of Cellulite 4BioSpecifics Technologies Corp. Announces Positive Data from Phase 2a Study of CCH for Treatment of Cellulite 5Global Hormone Replacement Therapy Market 2014-2018: Key Vendors are Actavis, Bayer, Merck & Co, Novartis and Novo Nordisk 2Science Power Jeffrey Epstein and Harvard University Fund Project to Visually Stop Tumor Growth 2Ontotext Releases Text Mining & Semantic Technology Running in the Cloud – Welcome to “S4? 2Ontotext Releases Text Mining & Semantic Technology Running in the Cloud – Welcome to “S4? 3Ontotext Releases Text Mining & Semantic Technology Running in the Cloud – Welcome to “S4? 4
... Iceland, and ALAMEDA, California, April 23, deCODE genetics ... CRA ) today announced the signing of agreements ... to deCODE,s,genetic markers for increased risk of major ... fibrillation (AF) and type 2,diabetes (T2D). These markers ...
... April 23 Cord Blood America, Inc. (OTC Bulletin ... company ( http://www.cordblood-america.com ) focused ... to families nationwide and internationally, which recently announced it ... said today it has eliminated another $817,000 of long-term ...
... honors President Carter, Carter Center for fight against neglected ... Progress (GHP) and ONE, as well as members of ... foundations and the business community, welcomed a delegation of ... reception on Capitol Hill today. The delegation presented a ...
Cached Biology Technology:deCODE and Celera Partner to Expand use of deCODE Risk Markers for Heart Attack, Stroke and Diabetes 2deCODE and Celera Partner to Expand use of deCODE Risk Markers for Heart Attack, Stroke and Diabetes 3deCODE and Celera Partner to Expand use of deCODE Risk Markers for Heart Attack, Stroke and Diabetes 4deCODE and Celera Partner to Expand use of deCODE Risk Markers for Heart Attack, Stroke and Diabetes 5deCODE and Celera Partner to Expand use of deCODE Risk Markers for Heart Attack, Stroke and Diabetes 6deCODE and Celera Partner to Expand use of deCODE Risk Markers for Heart Attack, Stroke and Diabetes 7Cord Blood America Further Reduction, $817,000, in Long-Term Debt 2African Health Delegates Welcomed to Washington 2
(Date:8/21/2014)... Fla. , Aug. 21, 2014 /PRNewswire-iReach/ ... that it has agreed to a partnership ... Orlando,s fastest growing Certified Nursing ... Gabriel Health Institute  an exclusive member of ... result, Binary Biometrics will improve its service ...
(Date:8/21/2014)... Nine researchers from Beth Israel Deaconess Medical Center ... "The World,s Most Influential Scientific Minds 2014," a ... ScienceWatch, a web resource for science metrics and research ... among the most influential "are performing and publishing work ... of their science," according to a Thomson Reuters statement. ...
(Date:8/21/2014)... a key region in the global oceanic circulation. "On the ... Atlantic water flows to the north into the Arctic Ocean ... sea ice push their way out of the Arctic into ... cools here on its way to the north and sinks ... this manner drives the global band of oceanic currents like ...
Breaking Biology News(10 mins):LiveScan Provider Binary Biometrics Announces Partnership with Preeminent CNA School, Gabriel Health Institute to Expand Services in Orlando Region 2LiveScan Provider Binary Biometrics Announces Partnership with Preeminent CNA School, Gabriel Health Institute to Expand Services in Orlando Region 3BIDMC researchers named among 'the most influential scientific minds' 2BIDMC researchers named among 'the most influential scientific minds' 3Arctic sea ice influenced force of the Gulf Stream 2Arctic sea ice influenced force of the Gulf Stream 3
... filled with widespread species, while moderate and higher elevations were ... is turning out to be partially incorrect. Widespread species now ... new study describes four species once considered to be the ... Plica plica . The study was published in ...
... researchers found that chicken eggs can provide a better ... professor in Clemson,s College of Agriculture, Forestry and Life ... Research Excellence award that allowed researchers to identify two ... congenital deafness and spine deformities in humans. Chapman ...
... turns out that your messenger RNA may catch more than one ... mRNA may travel one way down a cell, then board another, ... get where it ultimately needs to be. It,s a ... differentiating a brain cell from, say, a muscle cell, said ...
Cached Biology News:The collared treerunner is more than a single species 2Clemson research finds chickens offer clues to human birth defects 22-way traffic enable proteins to get where needed, avoid disease 2
...
...
Kit for testing the ability of a compound to simulate lipolysis in cultured human adipocytes. Kit contains controls, buffers and reagents for detecting glycerol and non-esterified free fatty acids. A...
The strip cap tool is a used for properly seating 8- and 12-cap strips in microplates or strip tubes. A grooved channel on one side is designed to seat domed caps, while the flat edge on the opposite...
Biology Products: