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Split-BioID: An Improved Method for Studying Protein-Protein Interactions

Posted by Beth Kenkel on May 1, 2018 9:57:26 AM

One way to define a protein’s purpose is by its protein-protein interactions (PPIs). These interactions are often modeled as binary relationships, i.e. protein A interacts with protein B; but proteins are social biomolecules. They can be part of multiple dynamic and overlapping complexes that have distinct functions. Many existing methods for identifying PPIs, such as affinity purification mass spectrometry (AP-MS), lack the ability to specifically identify proteins that interact with a particular protein  complex as opposed to an individual protein. The Bethune Lab has overcome this limitation by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent biotinylation technique BioID. The key advantage of Split-BioID is that it allows for the validation of a binary PPI as well as the identification of additional interacting factors.

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Topics: Plasmid Technology, Hot Plasmids

Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

Posted by Various Addgenies on Apr 23, 2018 10:00:31 AM

Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need. Below you'll find our hot plasmid articles from April 2018. If you'd ever like to write about a recent plasmid deposit please sign up here.

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Topics: Hot Plasmids, Podcast

AAVs for Genome Editing

Posted by Tyler Ford on Mar 27, 2018 9:32:43 AM

Guest blogger Todd Waldman, Professor at Georgetown University, contributed to this post.

Adeno-associated viruses (AAVs) make fantastic gene delivery vehicles for episomal gene expression and are particularly useful for gene delivery to the nervous system. For many years they have also been used to enhance the efficiency of genome editing. In this post we'll walk through a variety of ways you can use AAVs to improve your genome editing experiments (with and without targeted nucleases).

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Topics: Hot Plasmids, Viral Vectors

New Neuroscience Tool: The SF-iGluSnFr Glutamate Sensor

Posted by Tyler Ford on Mar 8, 2018 9:00:00 AM

In a previous blog post we discussed how fluorescent proteins can be used to construct biosensors, biological tools that monitor processes or detect molecules. Here we’ll be diving into the details surrounding SF-iGluSnFr, a recently upgraded biosensor designed to detect glutamate.

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Topics: Hot Plasmids, Fluorescent Proteins

Plasmids 101: Secondary Nanobody Toolbox

Posted by Beth Kenkel on Feb 27, 2018 9:04:41 AM

Western blots. ELISAs. Immunofluorescence. What do all of these techniques have in common? They all typically require secondary antibodies, frequently of the mouse or rabbit variety. While antibodies certainly aren’t “broken,” their production does require continued animal sacrifice. Could there be an alternative method for immunodetection? Enter the Görlich lab and their anti-mouse and -rabbit IgG secondary nanobodies toolbox. Nanobodies are like tiny antibodies which work just as well, if not better, than antibodies for all of the above listed molecular techniques, but they can also be expressed in bacteria and extracted with common protein purification methods. Read on to learn more about nanobodies and how their structure and function compare to IgG antibodies, as well as how to produce them for use in your lab.

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Topics: Hot Plasmids, Plasmids 101

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