Sep 01, 2022
Overall protocol for MicroPOTS LCMS top down proteomics of pancreas tissue sections
- 1Pacific Northwest National Laboratory
Protocol Citation: Mowei Zhou, James M Fulcher, Yen-Chen Liao, Ljiljana.PasaTolic 2022. Overall protocol for MicroPOTS LCMS top down proteomics of pancreas tissue sections. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l61oy5vqe/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 19, 2022
Last Modified: September 01, 2022
Protocol Integer ID: 68924
Abstract
This is the overall workflow for LCMS top down proteomics of pancreas functional units from tissue sections using the MicroPOTS platform. The expected outcomes are proteoform identification and quantitation values from selected tissue functional units.
Tissue collection
Tissue collection
The tissue sections were prepared and shipped from Vanderbilt-TMC following the protocol below:
Protocol
NAME
Cryostat Sectioning of Tissues for 3D Multimodal Molecular Imaging
CREATED BY
Jamie Allen
Sample preparation
Sample preparation
Functional units (islet and acinar) were dissected and collected into the microPOTS platform using the method below:
Protocol
NAME
Laser Capture Microdissection of Tissue Functional Units for microPOTS Top-Down ProteomicsCREATED BY
James M Fulcher
Data Acquisition
Data Acquisition
The samples were analyzed by LCMS top down proteomics as described below:
Protocol
NAME
Top Down Proteomics Data Collection for Microdissected Pancreas Tissue Functional UnitsCREATED BY
Mowei Zhou
Data Analysis
Data Analysis
LCMS datasets were analyzed for proteoform identification and quantitation. The final results are reported.
Protocol
NAME
Proteoform Identification and Quantitation with TopPIC and TDPortal for Human TissuesCREATED BY
Mowei Zhou