High pH fractionation

The recently acquired Vanquish Neo UHPLC system is ready to serve our customers. Optimized offline pre-fractionation at high pH allows quantifying 50.000 peptides per sample and outperforms traditional low pH offline fractionation. We are happy to offer a new SOP for deep proteome and phosphoproteome analysis for the samples with limited availability. Briefly, 50µg of HEK whole cell lysate was digested using the SP3 method and separated at pH 10 using C18 RP HPLC with 1µl/min flow rate. Every minute 1µl fraction is collected. Fractions are concatenated and analyzed by low pH HPLC-MS.