Phospho explorer antibody assay

Protein samples from ApoE treated islets and controls were used for the Phospho Explorer Antibody Assay (Full Moon BioSystems, CA). This assay performed a quantitative protein phosphorylation profiling of over 30 signaling pathways for control and ApoE treated islets. A list of upregulated phosphorylated proteins was obtained.

The Phospho Explorer Antibody Assay (Cat# PEX100, Fullmoon Biosystems, Sunnyvale, CA, USA) was used to measure phosphorylation status in EV- or TRIM22-transfected cells. This array was consisted of 1318 antibodies associated with various signaling pathways. Transfected cells were lysed with non-denaturing lysis buffer. Extracted proteins were biotinylated and then incubated on antibody array slides. After incubation, signals of proteins were detected by dye-labeled streptavidin. Raw signal intensity was normalized to all signals on the array slides. Fold changes between samples were calculated and proteins with fold changes above 1.2-fold and below 0.8-fold were included in the final dataset. Array experiments and analysis were performed as services by Ebiogen lnc.