Project description:This project aims at providing a quantitative dataset from LC-MS/MS injections of a calibrated UPS1 mixture spiked in an Arabidopsis thaliana background.

2022-07-31 | PXD027800 | Pride

Absolute quantification of mTOR related signaling molecules

Project description:To determine protein concentration in cells, we performed spike-in DIA experiments. Isotopically labeled concatemers (for mTOR-related signaling proteins) were spiked into lysates from p18Rev cells (each 20 mg) with four three-fold serial dilutions from 5,000 fmol. The lysates were subjected to digestion with trypsin. For absolute quantification of proteins, the digests containing isotopically labeled recombinant proteins were separated on a column (EV1106, Evosep) using a pre-programmed gradient (15 samples per day) at a flow rate of 200 nL/min using the Evosep One system (Evosep).

2024-05-23 | PXD042413 | JPOST Repository

in Vitro Brominated Peptides Spiked into U373 cell digestion

Project description:We performed nanoLC-MS/MS analysis of an in vitro generated, trypsin-digested brominated human serum albumin standard, spiked into a complex trypsin-digested proteomic background, in an LTQ-Orbitrap instrument. We found that brominated peptides spiked in at a 1-10% ratio (mass:mass) were easily identified by manual inspection when higher-energy collisional dissociation (HCD) and collision induced dissociation (CID) were employed as the dissociation mode; however, confident assignment of brominated peptides from protein database searches required a novel approach. By addition of a custom modification, corresponding to the substitution of a single bromine with 81Br rather than 79Br for dibromotyrosine (79Br81BrY), the number of validated assignments for peptides containing dibromotyrosine increased significantly when analyzing both high resolution and low resolution MS/MS data.

2013-08-28 | PXD000430 | Pride

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