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The Fluorescent Vegetables in Aptamer Soup

Posted by Aliyah Weinstein on Jan 29, 2019 9:27:00 AM

If you’re been following Addgene on #souptwitter or if you just enjoy a warm meal on a winter day, you should be happy to learn that many DNA-based reagents are named for soup ingredients! From pSOUP to mCherry, satisfying ingredients reagents permeate the molecular biologist’s kitchen lab - and scientists using aptamers might know this best, as most fluorescent aptamers are named for a fruit or vegetable!

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Topics: Fluorescent Proteins

Tetbow: Bright Multicolor Labeling for Neuronal Tracing

Posted by Guest Blogger on Jan 24, 2019 9:24:20 AM

This post was contributed by Richi Sakaguchi from Kyoto University, and Marcus N. Leiwe and Takeshi Imai from Kyushu University.

Stochastic multicolor labeling is a powerful solution for discriminating between neurons for light microscopy-based neuronal reconstruction. To achieve stochastic multicolor labeling, Brainbow used the Cre-loxP system to express one of the three fluorescent protein (XFP) genes in a transgene. When multiple copies of the transgene cassette are introduced, stochasticity will result in a combinatorial expression of these three genes with different copy numbers, producing dozens of color hues (Livet et al., 2007; Cai et al., 2013). However, the brightness of Brainbow was inherently low. This is because the stochastic and combinatorial expression of fluorescent proteins is only possible at low copy number ranges, resulting in low fluorescent protein level.

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Topics: Fluorescent Proteins

Fluorescent Biosensors for Measuring Autophagic Flux

Posted by Beth Kenkel on Jan 22, 2019 9:41:01 AM

Autophagy (Greek for “self-eating”) is a process by which cytoplasmic material, including organelles, are targeted to lysosomes for degradation. Autophagy is a dynamic process which involves autophagosome synthesis, delivery of materials to be degraded to the lysosome, and degradation of autophagic substrates inside the lysosome. Historically, methods for studying autophagy focused on counting the number of autophagosomes. This approach, however, has inherent limitations because it turns a dynamic process into a static measurement and it provides limited information about what materials or organelles are being targeted for autophagy. The development of several fluorescent autophagy reporters now allows for the measurement of autophagic flux, or the changes in autophagic activity, and are a more reliable indicator of autophagic activity. The aim of this post is to provide an overview of four autophagy biosensors currently available from Addgene.

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Topics: Plasmid Technology, Fluorescent Proteins

2019 Michael Davidson and Roger Tsien Commemorative Travel Awards

Posted by Jennifer Tsang on Jan 7, 2019 10:05:42 AM

UPDATE (06/04/2019): THE TRAVEL AWARD IS NOW CLOSED. READ ABOUT THE AWARDEES HERE.

Are you a masters student, PhD student, or postdoc using fluorescent proteins in your research? Consider applying for the Michael Davidson and Roger Tsien Commemorative Travel Awards.

To commemorate their innumerable contributions to the development of fluorescent protein tools, biological research, and their dedication to scientific sharing, Addgene is once again seeking 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 the next year (April 2019 - March 2020) who can demonstrate that fluorescent proteins have or will have an impact on their research.

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Topics: Fluorescent Proteins

When GFP lets you down

Posted by Guest Blogger on Aug 23, 2018 8:05:04 AM

This post was contributed by guest blogger Joachim Goedart, an assistant professor at the Section of Molecular Cytology and van Leeuwenhoek Centre for Advanced Microscopy (University of Amsterdam).

GFP is the most popular, most widely used genetically encoded fluorescent probe. Several factors contribute to the popularity of GFP including (i) fast and complete maturation to functional, fluorescent protein in almost all organisms and cell types, (ii) no need to add a co-factor, (iii) easy visualization with standard filter sets on a fluorescence microscope, and finally (iv) good toleration in fusion proteins.

Since GFP is such a well-validated, all-round good performing probe, it is the first choice when selecting a genetically encoded fluorescent tag. There are, however, a number of limitations that you may run into if you choose to use it. Several of these limitations and possible solutions are discussed below.

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Topics: Fluorescent Proteins

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