• [User Manual] KILR® Retroparticles For Cytotoxicity Assays

  The KILR Retroparticles provide a potent vehicle for the stable delivery of the KILR reporter construct into almost any dividing cell line, enabling the creation of KILR cytotoxicity assays in the cell line expressing the target of your choice. The KILR Cytotoxicity cell line you generate enables the development of a simple, non-radioactive and dye-free method ...

  

Assay Principle

The KILR retroviral particle is a Moloney Murine Leukemia Virus (MMLV) engineered to drive expression of the KILR reporter construct, a housekeeping gene tagged with Enhanced ProLabel® (ePL), a β-galactosidase (β-gal) reporter fragment. The KILR Retroparticles for Suspension cells contain a hybrid LTR promoter for expression of the fusion protein in difficult-to-transduce suspension cell lines, whereas the KILR Retroparticles for Adherent cells use a CMV promoter for expression of the KILR construct in most adherent cell lines. When stably transduced with the KILR Retroparticles, the target cells express the KILR reporter construct inside the cells. Death of the cells leads to rupture of the cell membrane and the KILR reporter protein is released into the medium. Addition of KILR detection reagent, containing the complementing β-gal reporter fragment, Enzyme Acceptor (EA), results in complementation of the two enzyme fragments (EA and ePL; See Figure 1). The resulting active enzyme hydrolyzes substrate to generate a chemiluminescent signal.

Applications of the KILR Cytotoxicity Assays