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        • Stable Cell Lines for Drug Screening in CNS

ALS Model: FUS-TLS Stress Granules Assay Cell Line

Reference: P30716


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Data Sheet

 
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder that is characterized by premature degeneration of motor neurons, resulting in a progressive, fatal paralysis. Basis of this in vitro model is that ALS-linked fused in sarcoma/translocated in liposarcoma (FUS/TLS or FUS) is concentrated within cytoplasmic stress granules under conditions of induced stress. A stable cell line expressing fluorescent FUS/TLS has been developed and validated to form stress granules under under conditions of oxidative stress.

Innoprot offers this cell line as a “stable cell line” but it is also offered as vials of division-arrested cells (DA cells) in order to perform a small number of assays. Each vial of these DA cells contains 2 million cells, an enough number of cells to perform a complete experiment in a 96 well-plate.
 


Assay Details


U2O2 cells stably expressing human FUS/TLS were induced with IPTG 5mM during 48h to produce the hFUS-tGFP protein. Subsequently, cells were incubated with the compounds at 10 uM during 24 hours and then, cells were treated with 300 uM sodium arsenite  during 120 min to induce the stress granules formation. The FUS/TLS-tGFP fluorescent granules  were quantified using the BD Pathway HCS Reader and Attovision Compartimentalization Software.

Results indicate that the detecting dynamyc range is dependent on the inhibitor biophysics and biochemical characteristics and the treatment time. This stress granules assay was validated with an average of Z´= 0.62+/- 0.01 for High Content Screening with a 24 hours treatment.
  • (More information and results: see data sheet or request us the poster of the product)


Applications

  • The stably transfected FUS/TLS cell line can be used in drug discovery for pathological stress granules formation inhibitors.
  • This model permits to evaluate the FUS/TLS protein distribution in living cells studying the protein localization pattern in the space and time.
  • This model provides a strategy to evaluate drugs that not have cell permeability.
  • This cellular model have been adapted to HCS analyses based on image algorithms to test cytosolic and nuclear globs generation process.
    

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