Reverse Phase Protein Arrays

Reverse Phase Protein Arrays (RPPA) represent a sensitive antibody based proteomic approach, which enables simultaneous quantification of numerous proteins and post-translational modifications across a multiple sample set. Combining the speed and accuracy of the ArrayJet Mercury 100 slide printer with the scanning power of the infra-red Innopsys Innoscan 710 allows us to profile pathways rapidly with high sensitivity.

• Dot-blot immunoassay that utilises miniaturised arrays to measure protein expression in biological samples
• Fully customisable from a validated portfolio of >250 antibodies covering key cancer survival, cell–cycle and canonical signalling pathways
• Samples per run: 12 or 36 samples
• Measure up to 120 antibodies
• Requirements: Protein lysate at a minimum of 2 mg/ml (50 µl)
• Contact rppa@igc.ed.ac.uk
• Further information

Pathway profiling of a novel SRC inhibitor, AZD0424, in combination with MEK inhibitors for cancer treatment (2022). John C Dawson , Alison Munro, Kenneth Macleod, Morwenna Muir, Paul Timpson, Robert J Williams, Margaret Frame, Valerie G Brunton, Neil O Carragher.

Pathway profiling of a novel SRC inhibitor, AZD0424, in combination with MEK inhibitors for cancer treatment - PubMed

Colleagues at the Institute of Genetics & Cancer used RPPA to profile changes in intracellular signalling pathways following exposure to SRC inhibitor AZD0424 alone and in combination.

Using RPPA they showed elevation of SRC following MEK inhibition across a panel of breast cancer cell lines, together with a concentration-dependent decrease in SCR-family kinase activation. A reduction in phosphorylation of the SRC kinase target STAT5 (Tyr694) was also noted, in addition to EGFR family (Tyr1248/Tyr1173), PLCγ (Tyr783) and SHP2 (Tyr542) signalling

As SRC inhibitors perform poorly as single anti-cancer agents in most cancer, Dawson et al sought to identify potential resistance mechanisms that rely upon SRC that could be targeted using AZD0424. Using colorectal cell lines with mutant KRAS, they showed treatment with MEK inhbitors trametinib or AZD6244 induced activation of SRC, and in addition compensatory induction in phosphorylation of a number of key proteins involved in EGFR. Cells treated with a combination of AZD0424 and a MEK inhibitor blocked the activation of EGFR, SHP2, PLCγ suggesting that reactivation of signalling through the EGFR pathway in response to MEK inhibition requires SRC activity.