SEAP Chemiluminescence Kit 2.0

Chemiluminescent detection of protein-protein interactions

Chemiluminescent detection of protein-protein interactions. The bait vector pM-53 expresses p53 fused to the GAL4 DNA-binding domain. When HEK 293 cells were cotransfected with this bait together with the SV40 large T antigen prey vector, pVP16-T, strong expression of SEAP was detected because large T antigen interacts with p53. The CP protein, which does not interact strongly with p53, was used as a negative control.

SEAP activity is highly specific, with a >20-fold increase in activity

Comparison of the sensitivities of SEAP and firefly luciferase. Parallel cultures of BHK cells were transiently transfected with the indicated amounts of either pSEAP2-Control Vector or a similar vector expressing luciferase. After 24 hr, SEAP activity was assayed in the appropriate culture media using the Great EscAPe chemiluminescent assay. Similarly, after 24 hr, cell lysates were prepared from the luciferase cultures, and luciferase activity was assayed using a commercial kit. RLU = relative light units.

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription.