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Fuse-It-P

Fuse-It-P View larger

A fusion reagent used to transfer water-soluble proteins into the cytoplasm of living cells

  • The lumen of fusogenic liposomes is used as a carrier for water-soluble proteins
  • Superior fusion efficiency (up to 80-100%)
  • Fusion process completed within 1 - 20 minutes
  • Optimized for the transfer of low and intermediate protein amounts to prevent concentration-induced artifacts in cell behavior
  • Lyophilized

in cooperation with

logo_beniag.jpg

infrared fluorescent

Product Details

Applications:

  • Transfer of labeled proteins or peptides into living cells for functional imaging, speckle analysis, FRAP, or single molecule analysis, etc.
  • Incorporation of antibodies or blocking antibodies into living cells
  • Blocking, induction, or replacement of protein-induced/regulated signal cascades
  • Re-incorporation of proteins into mutant cells

Transferred proteins are instantly active inside the cells, and after membrane fusion, cells can immediately be used for further analysis. Since proteins are freed directly into the cytoplasm, no partial or complete lysosomal degradation can occur, as is typical for other endosomal uptake-depending proteofection methods. Depending on your experimental needs, the fusion process can be monitored by fluorescence microscopy.

Specifications:

Form Lyophilized powder
Storage -20 °C
ExMax/EmMax 750/780 nm (infrared)


NOTE
: Transfer of heavily charged proteins can affect fusion efficiency.

Glycerol (>5 %) and BSA will obstruct the transport of peptides/proteins.

Fuse-It-P vesicles were filled with LifeAct and fused with myofibroblasts. A bright staining of the actin cytoskeleton can be observed after 5 minutes.

For details please watch the according videos:

Individual Staining

Overlay

Principle of Membrane Fusion

The incorporation of small liposomal carriers into the plasma membrane of mammalian cells is the idea behind all of ibidi’s Fuse-It products. Liposomal carriers are able to attach and instantly fuse with plasma membranes in a physicochemical-driven manner. ibidi’s new Fuse-It reagents efficiently use this mechanism and fuse with mammalian cell surfaces immediately upon contact. Therefore, this novel technique makes the transfer of molecules independent of biological processes, such as endocytosis, pinocytosis, or specific receptor binding.

Supporting Material

Movies

User Comments

Stephane R. Gross, Aston University, Birmingham, United Kingdom

“For my group, the efficient labeling of cells is an extremely important issue. Since most of our cells are very sensitive to changing medium conditions, mild and fast labeling is a must. We have found, in the very short time we have used the system and therefore without real optimization, that both the Fuse-It-Color and Fuse-It-P were very well functional, with the latter perfectly capable of delivering our target proteins into HeLa cells. Such delivery was obtained using relatively low amounts of exogenous proteins and incubation times as short as 5 minutes.”

Dr. Stephane R. Gross
Lecturer in Cellular Biology
School
of Life and Health Sciences
Aston
University Birmingham
United Kingdom

http://www.aston.ac.uk/lhs/research/


Michael Börsch, Jena University Hospital, Friedrich Schiller University, Jena, Germany

"The incorporation of transmembrane proteins from proteoliposomes or detergent-containing buffers into black lipid membranes is a very dicey issue. I have used your high fusogenic Fuse-It-P for this purpose and it really worked perfectly. It was an extremely simple preparation, and the incorporation was very reliable and easily adjustable. It made this very difficult step for all experiments in my institute much easier. Thanks a lot."

Prof. Dr. Michael Börsch
Single-Molecule Microscopy Group
Jena University Hospital
Friedrich Schiller University Jena
Germany

http://www.m-boersch.org/index.html

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