Project description:We performed an RNA Sequencing experiment on dorsal hippocampal tissue from four groups of animals: Baf53b+/- homecage (Baf53b+/- HC); Baf53b+/- behavior (Baf53b+/- Beh); wildtype homecage (WT HC); and wildtype behavior (WT Beh). Homecage animals were sacrificed directly from the animal's cage. Behavior animals were sacrificed thirty minutes following Object Location Memory training. The objective of this study was to examine activity regulated gene expression following a learning event (HC vs Beh) in wildtype and Baf53b+/- mutant mice.
Project description:40 µL plasma from a pool of 20 neurologically healthy donors was separated into a bound and a depleted fraction using a MARS Hu-14 column. This experiment represents the bound fraction. Sample was in-solution trypsin digested and peptides were fractionated into 10 fractions by mixed-mode reversed phase anion exchange (MM(RP-AX) using a Promix MP column. Each fraction was analyzed separately by LC-MS/MS on an Orbitrap Velos Pro mass spectrometer. Resulting data was searched using SeachGui, summarized in PeptideShaker and exported to the CSF-PR.
Project description:40 µL plasma from a pool of 20 neurologically healthy donors was separated into a bound and a depleted fraction using a MARS Hu-14 column. This experiment represents the depleted fraction. Sample was in-solution trypsin digested and peptides were fractionated into 67 fractions by mixed-mode reversed phase anion exchange (MM(RP-AX) using a Promix MP column. Each fraction was analyzed separately by LC-MS/MS on an Orbitrap Velos Pro mass spectrometer. Resulting data was searched using SeachGui, summarized in PeptideShaker and exported to the CSF-PR.
Project description:1.6 mL CSF from a pool of 21 neurologically healthy donors was separated into a bound and a depleted fraction using a MARS Hu-14 column. This experiment represents the bound fraction. Sample was in-solution trypsin digested and peptides were fractionated into 10 fractions by mixed-mode reversed phase anion exchange (MM(RP-AX) using a Promix MP column. Each fraction was analyzed separately by LC-MS/MS on an Orbitrap Velos Pro mass spectrometer. Resulting data was searched using SeachGui, summarized in PeptideShaker and exported to the CSF-PR.
Project description:1.6 mL CSF from a pool of 21 neurologically healthy donors was separated into a bound and a depleted fraction using a MARS Hu-14 column. This experiment represents the depleted fraction. Sample was in-solution trypsin digested and resulting peptides were fractionated into 66 fractions by mixed-mode reversed phase anion exchange (MM(RP-AX) using a Promix MP column. Each fraction was analyzed separately by LC-MS/MS on an Orbitrap Velos Pro mass spectrometer. Resulting data was searched using SeachGui, summarized in PeptideShaker and exported to the CSF-PR.
Project description:We compared the performance of multidimensional protein identification (MudPIT) on Velos Pro Orbitrap (VPO) and Velos Orbitrap Elite (VOE) mass spectrometers to single dimension reversed phase (RP) chromatography on a Q Exactive Plus (QE+) and an Orbitrap Fusion™ Lumos™ (OFL). Using a digested HeLa cell protein extract, we carried out 16 different chromatography conditions on four different instrumentation platforms with three replicates of each condition, for a total of 48 proteomics analyses. We first compared selected chromatography conditions of the QE+ and OFL by varying column lengths, inner diameters, and C18-RP particle sizes. We next selected one chromatography condition on each system and varied the effective RP gradients lengths