ADP-Glo™ Lipid Kinase Assay Technical Manual

Instructions for Use of Product(s)
V1711, V1701, V1721, V1731, V1741, V1751, V1761, V1771, V1781, V1782, V1791, V1792, V1690, V1691: PI3K Class I Enzyme System

Literature # TM365

ADP-Glo™ Lipid Kinase Systems provide a complete set of reagents for performing phosphoinositide lipid kinase (PIK) reactions using a luminescent ADP-detection platform, the ADP-Glo™ Kinase Assay. The systems include purified recombinant proteins of class I PI3Ks, optimized reaction buffer and ready-to-use lipid kinase substrates. The lipid kinase reaction is performed by incubating lipid substrate with a recombinant enzyme and ATP, and the kinase activity is measured using the ADP-Glo™ Kinase Assay. The ADP-Glo™ Kinase Assay is performed in two steps. First, after the kinase reaction, an ATP-depletion reagent is added to terminate the lipid kinase reaction and deplete any remaining ATP, leaving only ADP. Second, a detection reagent is added to simultaneously convert ADP to ATP and allow the newly synthesized ATP to be converted to light using a coupled luciferase/luciferin reaction.

Summary of Changes
The following changes were made to the 10/22 revision of this document:
1. Corrected protocol step numbering in Section 3.B.
2. Updated disclaimers.
3. Made minor text edits.
4. Updated cover page and document font.

Revised 10/22.