Sep 01, 2022
Overall protocol for MicroPOTS LCMS top down proteomics of kidney tissue sections
- 1Pacific Northwest National Laboratory
- Human BioMolecular Atlas Program (HuBMAP) Method Development Community
- PNNL-TTD
Protocol Citation: Mowei Zhou, James M Fulcher, Yen-Chen Liao, Ljiljana.PasaTolic 2022. Overall protocol for MicroPOTS LCMS top down proteomics of kidney tissue sections. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2lynm1qvx9/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: 68871
Funders Acknowledgements:
National Institutes of Health (NIH) Common Fund, Human Biomolecular Atlas Program (HuBMAP)
Grant ID: UG3CA256959-01
Abstract
This is the overall workflow for LCMS top down proteomics of kidney 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:
Sample preparation
Sample preparation
Functional units (glomerulus, medullary, tubule) were dissected and collected into the microPOTS platform using the method below:
Protocol
CREATED BY
James M Fulcher
Data Acquisition
Data Acquisition
The samples were analyzed by LCMS top down proteomics as described below:
Protocol
CREATED BY
Mowei Zhou
Data Analysis
Data Analysis
LCMS datasets were analyzed for proteoform identification and quantitation. The final results are reported.
Protocol
CREATED BY
Mowei Zhou