AKT Multiplex Immunoassay (Panels 1-3)
ERK/MEK Multiplex Immunoassay (Panels 4-5)

The Signaling Multiplex Immunoassays are novel isoform specific signaling multiplex immunoassay panels of 12 biomarkers for AKT and rpS6 and 8 biomarkers for MEK and ERK to measure on target or downstream pathway modulation by multiple classes of drugs including PI3K K/AKT/ mTORC and RAS/RAF/MEK/ERK inhibitors The AKT/rpS6 multiplex immunoassays were built on the Luminex platform by grouping 12 biomarkers into three panels Signaling Multiplex Panel 1 ("Total AKT/rpS6 Panel") detects total levels of AKT1, AKT2, AKT3, and rpS6 Signaling Multiplex Panel 2 ("Phospho AKT/rpS6 A Panel") detects pSer473-AKT1, pSer474-AKT2, pSer472-AKT3, and pSer235-rpS6 and Signaling Multiplex Panel 3 ("Phospho AKT/rpS6 B Panel") detects pThr308-AKT1, pThr309-AKT2, pThr305-AKT3, and pSer240/244-rpS6. The MEK/ERK multiplex immunoassays were built on the Luminex platform by grouping 8 biomarkers into two panels Signaling Multiplex Panel 4 ("Total MEK/ERK Panel") measures total levels of ERK1, ERK2, MEK1, and MEK2; Signaling Multiplex Panel 5 ("Phospho MEK/ERK Panel") measures pThr202/Y204-ERK1, pThr185/Y187-ERK2, pSer218/222-MEK1, and pSer222/226-MEK2. Total lysates from a single core needle biopsy is sufficient to evaluate all five multiplexes.

1. Specimen Collection
   • Shipment to PADIS, FNLCR
   • Shipment to EET Biobank
2. Sample Preparation
3. Kit Use and Data Reporting SOP
   • AKT/rpS6
   • MEK/ERK

• Assay Training and Key Reagent Support
• NCI Publications