Cell Line Stability 

Cells will undergo genotypic changes over time resulting in reduced responsiveness in normal cell culture conditions. Genetic instability is a biological phenomenon that occurs in all stably transfected cells. Therefore, it is critical to prepare an adequate number of frozen stocks at early passages. HEK-Blue™ KO-ALPK1 cells should not be passaged more than 20 times to remain fully functional.

HEK-Blue™ KO-ALPK1 cells were generated from HEK-Blue™ Null1-v cells through the stable knockout of the ALPK1 gene. The parental cells derive from human embryonic kidney 293 (HEK-293) cells. HEK-Blue™ KO-ALPK1 cells express a secreted embryonic alkaline phosphatase (SEAP) under the control of an NF-κB inducible promoter comprised of an IFN-β minimal promoter fused to five NF-κB and AP-1 binding sites. Levels of SEAP in the supernatant can be easily determined with HEK-Blue™ Detection, a SEAP detection cell culture medium. Unlike their parental cell line, HEK?Blue™ KO-ALPK1 cells do not respond to cytosolic ADP-Heptose. However, they do respond to other NF-κB inducers such as TNF-α and IL-1β. HEK?Blue™ KO-ALPK1 cells are resistant to Zeocin™