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Innoprot

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        • Stable Cell Lines for Drug Screening in CNS

ALS Model: TDP-43 Stress Granules Assay Cell Line

Reference: P30710


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Data Sheet



Amyotrophic lateral sclerosis (ALS) is one of the most common degenerative disease of the motor neuron system.The TDP-43 pathology seems to be a dominant type of pathology across sporadic ALS types.  Innoprot has developed a novel fluorescence cell-based assay for High Content Screening that allows the quantification of pathological TDP43 globs  into the nucleus and cytosol

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 TAR DNA-binding protein 43 (TDP-43) were induced with IPTG 5mM during 48h to produce the hTDP43-tGFP protein. Subsequently, cells were incubated with the compounds at 10 mM during 24 hours and then, cells were treated with 250 uM sodium arsenite  during 90 min to induce the stress granules formation. The TDP43-tGFP nuclear globs  were quantified using the BD Pathway HCS Reader and Attovision Compartimentalization Software. Error bars represent the standard deviation among 3 replicate wells.

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 TDP43 cell line can be used in drug discovery for pathological globs formation inhibitors.
  • This model permits to evaluate the TDP-43 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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