Michael Davidson and Roger Tsien Commemorative Travel Awards

Posted by Tyler Ford on Jan 5, 2017 10:55:23 AM

 

UPDATE (February 1, 2017): THE TRAVEL AWARD IS NOW CLOSED - AWARDEES HAVE BEEN NOTIFIED AND WILL BE FEATURED IN A FUTURE POST

To commemorate their innumerable contributions to the development and use of fluorescent protein tools and their dedication to scientific sharing, Addgene is opening applications for the Michael Davidson and Roger Tsien Commemorative Travel Awards. These $2,000 USD awards will be open to any masters students, PhD students, or postdocs traveling to an academic conference in 2017 who can demonstrate that fluorescent proteins have or will have an impact on their research.

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Topics: Fluorescent Proteins

Better Dyeing Through Chemistry & Small Molecule Fluorophores

Posted by Guest Blogger on Sep 8, 2016 10:30:00 AM

This post was contributed by guest blogger, Luke Lavis, a Group Leader at the Janelia Research Campus, Howard Hughes Medical Institute.

Chemistry is Dead, Long Live Chemistry!

The discovery of green fluorescent protein (GFP) sparked a renaissance in biological imaging. Suddenly, cell biologists were no longer beholden to chemists and (expensive) synthetic fluorophores. Add a dash of DNA with an electrical jolt and cells become perfectly capable of synthesizing fluorophore fusions on their own. Subsequent advances in fluorescent proteins have replicated many of the properties once exclusive to small-molecules: red-shifted spectra, ion sensitivity, photoactivation, etc. These impressive advances lead to an obvious question: In this age of GFP and its ilk, why should cell biologists talk to chemists?

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Topics: Imaging, Fluorescent Proteins

Editor's Choice, July 2016

Posted by Tyler Ford on Aug 5, 2016 11:00:00 AM

To better highlight the great content contributed by our bloggers each and every month, we've decided to start an "Editor's Choice" series. Each month, I'll summarize the most popular post of the month and point out one or more additional posts that deserve a peek in case you missed them.

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Topics: CRISPR, Fluorescent Proteins, Editor's Choice

Special Delivery: Fluorophore Targeting for FRET Studies

Posted by Guest Blogger on Jul 19, 2016 10:30:00 AM

This post was contributed by guest blogger James D. Fessenden, an Assistant Professor at Brigham and Women’s Hospital.

Biochemists often struggle to understand how a protein of interest actually behaves. How large is it? What parts of it move when you feed it substrate or add an essential cofactor? How many binding partners does it have and how do they come off and on in a cellular environment? If these are pressing issues in your laboratory, then FRET experiments are a viable biophysical path to answers.

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Topics: Lab Tips, Fluorescent Proteins

When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

Posted by Guest Blogger on Apr 19, 2016 10:30:00 AM

This post was contributed by guest blogger Erik L. Snapp.

Stop using EGFP/GFP for fusion proteins! Despite multiple studies in high profile journal articles, many researchers remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant artifacts.

If you're using green fluorescent protein or Enhanced Green Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label of cells, there's no problem. You're OK. However, if you fuse your protein of interest (POI) to GFP to study the protein's behavior in cells, in solution or something in between, you are using a tag with a serious drawback. The standard EGFP plasmid that used to be sold by Clontech and is in a freezer box in just about every lab in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently dimerize. That means that your POI fused to GFP or another fluorescent protein (FP) could be forming dimers in cells. Why should you care? Three simple ways a dimeric FP could ruin your day (and experiment) are listed below. Solutions to avoid these all too common issues follow.

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Topics: Imaging, Fluorescent Proteins

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