Latest Posts

All Posts

Lighting Up Cell Signaling with Photoswitchable Kinases

Posted by Beth Kenkel on Jul 25, 2017 9:20:50 AM

Signal transduction pathways are a lot like cell phone networks. Protein kinases deliver messages to the next members of their pathways, but where the kinases are located, their signal strength, and how long their signals last all impact transduction of the message. To study signaling pathways, scientists frequently use growth factors or serum to stimulate a pathway of interest, but there can be a lot of static since other signaling networks can also be non-specifically activated and, like the game of telephone, often the signal must be transmitted by intermediate messengers. Optical control of kinase activity can provide greater spatiotemporal resolution than pharmacological or genetic approaches, but only a few such methods exist and they only work for a subset of kinases.

Read More >

Topics: Fluorescent Proteins

Plasmids 101: Introduction to FRET

Posted by Jason Niehaus on Jun 27, 2017 9:03:20 AM

Imagine being able to determine whether two proteins are within 10 nanometers of each other, or measure the tension in the helical structure of spider silk, or the activity of a protein in a synapse. What kinds of tools enable us to measure these properties, and what fascinating experiments could push these tools even further? All of these things can be done using FRET! Read on to find out more about this amazing imaging technique and find further tips for using FRET in your experiments here.

 

Read More >

Topics: Plasmids 101, Fluorescent Proteins

Plasmids 101: Visualizing Subcellular Structures & Organelles

Posted by Susanna Bachle on Jun 22, 2017 10:30:00 AM

The same way the human body is made up of organs, cells comprise compartments and structures, called organelles. Take a sneak peak inside a cell with the images from the Allen Cell Explorer (1).

When studying the function of a protein or its role in a disease, researchers often isolate proteins of interest and examine them using biochemical methods thus removing the context of the cell. However, much knowledge about functionality can be gained by understanding the location and transport of the protein within a living cell. Analyzing differences in protein localization and transport between healthy and diseased states can also provide interesting insights into disease mechanisms and protein function.

Read More >

Topics: Fluorescent Proteins

Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

Posted by Guest Blogger on Jun 15, 2017 10:30:00 AM

This post was contributed by guest bloggers Joachim Goedhart and Marieke Mastop from the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced microscopy, University of Amsterdam.

The previous two posts in this series described a practical approach to selecting a bright fluorescent protein and a photostable fluorescent protein. In the third post of this series, we will discuss how to select a non-aggregating fluorescent protein.

In the jellyfish Aequorea victoria, AvGFP forms a homodimer. In corals, the red fluorescent proteins form tetramers. In general, fluorescent proteins have a natural affinity and a tendency to form higher order aggregates. This property can be tolerated in some applications (e.g. labeling of cells or tracking promotor activity), but it is problematic in applications in which the fluorescent protein is used as an inert protein module. This is explained in more detail here. There are a variety of methods that can be used to measure your fluorescent protein’s propensity to aggregate. The basics and pitfalls of these experiments are discussed here.

Read More >

Topics: Fluorescent Proteins, Choosing the Brightest Fluorescent Protein

Choosing the B(right)est Fluorescent Protein: Photostability

Posted by Guest Blogger on Jun 8, 2017 10:30:00 AM

This post was contributed by guest bloggers Joachim Goedhart and Marieke Mastop from the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced microscopy, University of Amsterdam.

The previous post in this series described a practical approach to selecting a bright fluorescent protein. In the second post of this series, we will discuss how to select a photostable fluorescent protein.

Photobleaching is the irreversible destruction of a fluorophore under the influence of light. Any fluorescent molecule will photobleach at some point. For live-cell imaging, it is desirable to have fluorescent proteins that are photostable. On top of photobleaching, fluorescent proteins may display reversible intensity changes (Shaner et al, 2008; Bindels et al, 2017) and photoswitching (Kremers et al, 2009), which usually are undesired properties. In the ideal situation, a fluorescent proteis should emit a stable fluorescence signal, showing no or little deterioration or change of the signal during the course of the experiment.

Read More >

Topics: Fluorescent Proteins, Choosing the Brightest Fluorescent Protein

Blog Logo Vertical-01.png

Subscribe to Our Blog