Features & Solutions

Applications

 

Labeling in anaeroby

 

FAST is a user-friendly, real-time fluorescent reporting system working in low-oxygen or fully anaerobic conditions, opening great prospects for anaerobic biology.

You are interested in

visualizing protein localization?

characterizing gene-expression activity in Clostridium or other obligate anaerobes?

monitoring the dynamics of multi-species living biofilms in bioreactors?

Compatible techniques

MICROSCOPY

FLOW CYTOMETRY

MICROPLATE READERS

How can fast help you?

  • FAST is a small purely monomeric protein, of only 14 kDa. Its small size and minimal genetic footprint (375 bp) minimize the risk of perturbation linked to the use of genetic tags.
  • FAST binds non-covalently TFFluorogens, fluorogenic synthetic dyes that are dark in water and fluoresce only when bound to FAST, allowing specific detection of FAST.
  • FAST provide strong fluorescence immediately upon addition of TFFluorogens even in fully anaerobic conditions.

Other key features:

  • As TFFluorogen binds FAST non-covalently, labeling is non-permanent and can be easily reversed by washing TFFluorogens away.
  • The level of labeling can be controlled by adjusting the concentrations of TFFluorogens.
  • TFFluorogens come in a variety of colors: TFCoral (λem 600 nm), TFCitrus (λem 560 nm), TFAmber (λem 558 nm), TFLime (λem 540 nm).

recent papers about this application

A strongly fluorescing anaerobic reporter and protein-tagging system for Clostridium organisms based on the Fluorescence-Activating and Absorption-Shifting Tag (FAST) protein

Published in Appl. Environ. Microbiol. AEM-00622 (2019)

In this paper, Pr. Terry Papoutsakis from University of Delaware shows that FAST...

…associated with the fluorogenic ligands TFLime and TFCoral is highly fluorescent in Clostridium acetobutylicum in fully anaerobic conditions. His team gathered a comprehensive bunch of results, in flow cytometry or in fluorescent microplate readers, for FACSing or for visualizing protein localization, e.g., divisome, paving the way towards studies on “cell sorting, sporulation dynamics, and population characterization in pure as well as mixed cultures such as those in various native or synthetic microbiomes and syntrophic cultures”.

The inducible chemical-genetic fluorescent marker FAST outperforms classical fluorescent proteins in the quantitative reporting of bacterial biofilm dynamics.

Published in Sci. Rep. 8(1), 10336 (2018)

In this paper, Dr. Nelly Henry from Sorbonne Université (Paris, France) reports FAST...

…for the real-time quantitative analyses of the living community functions in a bacterial biofilm. Her team showed that GFP or mCherry fluorescence signal saturated after only a few hours of growth due to the reduction of oxygen concentration induced by bacterial consumption. In contrast, FAST allowed to increase the reporting dynamic range by two orders of magnitude, demonstrating its superiority for investigating dynamics in the complex environment of a living biofilm.

Other Applications

Labeling in anaeroby

Detecting protein-protein interactions

Protein & cell labeling

One tag, many colors

Reversible labeling, Silent Tag

A toolkit for sensing

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