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RANbodies: Reporter Nanobody Fusions

Posted by Beth Kenkel on Apr 10, 2018 8:56:44 AM

Antibodies are a go-to tool for detecting a protein of interest in cells and tissues. Although antibody production is well established, it’s also a process that’s difficult for individual labs to complete. The nanobody based RANbody platform from the Sanes Lab overcomes this limitation and allows for the flexible design and small scale production of antibodies.

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Topics: Plasmid Technology

CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

Posted by Guest Blogger on Feb 13, 2018 9:51:55 AM

This post was contributed by guest blogger Matthew J. Niederhuber, a graduate student at UNC Chapel Hill.

Chromatin immunoprecipitation followed by high-throughput sequencing, ChIP-Seq, is the go-to method for mapping where a protein binds genome-wide, and has been widely applied in many model organisms and cell lines. Although ChIP-seq is a relatively simple and robust protocol it does have limitations. The enzyme-based CUT&RUN method overcomes many of these limitations and makes it easier for you to map protein-DNA interaction with limited biological materials.

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Topics: Plasmid Technology, Techniques

In Vivo Biotinylation of Bacterial Fusion Proteins

Posted by Guest Blogger on Jan 25, 2018 9:09:35 AM

This post was contributed by guest blogger Jon Backstrom, a biochemist in the Vanderbilt Eye Institute and Tonia Rex's lab.

A common strategy to determine the binding kinetics of a purified protein involves immobilization on a solid support. This allows washing away of unbound material to calculate the amount of bound ligand (after subtracting out non-specific binding). Historically, glutathione-S-transferase (GST) fusion proteins have been immobilized on a reduced glutathione matrix. The advantage of a fusion protein is the efficient purification of an already immobilized target protein. The disadvantage is that the GST moiety, which forms dimers, may influence binding kinetics of the target ligand. Another important consideration is whether the affinity of an experimental protein-ligand interaction approaches that of GST-glutathione.

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Topics: Plasmid Technology, Hot Plasmids, Techniques

Top Requested AAV of 2017: pmSyn1-EBFP-CRE

Posted by Tyler Ford on Jan 17, 2018 9:57:12 AM

We began distributing ready-to-use virus preps through our viral service in late 2016 and requests are still pouring in! While our lentiviral service is going strong, the AAV service has shown incredible growth this year. pAAV-hSyn-DIO-hM4D(Gi)-mCherry was the top requested AAV prep for the 2nd year running, and you can learn more about this useful, DREADD-containing AAV here. But the top requested AAV that became available in 2017 is pmSyn1-EBFP-Cre from Hongkui Zeng’s lab. This AAV has had over 150 orders since coming online!

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Topics: Plasmid Technology, Hot Plasmids, Viral Vectors

Top Requested Plasmid of 2017 - pMD2.G

Posted by Tyler Ford on Jan 10, 2018 11:13:59 AM

Plasmid technologies are constantly evolving, but sometimes a technology is so useful it forever enhances biological research and discovery. CRISPR is a great example (the top requested plasmids from 2015 and 2016 were CRISPR plasmids), but so are lentiviral vectors, many of which are used to deliver Cas9 and other genes to mammalian cells. For this reason, the top requested plasmid of 2017 is the lentivirus envelope plasmid pMD2.G from Didier Trono’s lab!

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Topics: Plasmid Technology, Hot Plasmids, Viral Vectors

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