May 21, 2025
Plasma isolation from whole mouse
- Sierra Palumbos1,
- Erika Holzbaur1
- 1University of Pennsylvania
Protocol Citation: Sierra Palumbos, Erika Holzbaur 2025. Plasma isolation from whole mouse. protocols.io https://dx.doi.org/10.17504/protocols.io.3byl414zolo5/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: May 19, 2025
Last Modified: May 21, 2025
Protocol Integer ID: 218541
Keywords: plasma, plasma isolation of adult mice, plasma isolation from whole mouse protocol, plasma isolation, whole mouse protocol, adult mice, describing plasma isolation, mouse handling, euthanasia, isolation
Funders Acknowledgements:
National Institute of Neurological Disorders and Stroke
Grant ID: R01-NS060698
National Institute of Neurological Disorders and Stroke
Grant ID: F32-NS129586
Michael J. Fox Foundation for Parkinson's Research (MJFF)
Grant ID: MJFF-021130
Michael J. Fox Foundation for Parkinson's Research (MJFF)
Grant ID: MJFF-15100
Michael J. Fox Foundation for Parkinson's Research (MJFF)
Grant ID: MJFF-019411
Abstract
Protocol describing plasma isolation of adult mice. Used in Palumbos et al., 2025. Adapted from Goldsmith et al., 2022.
All mouse handling and euthanasia must follow IACUC regulations.
Guidelines
Prior approval from an Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee is required for this experiment.
Troubleshooting
Trunk blood collection
Animal is euthanized following IACUC protocol (e.g., isopropanol overdose)
Animal is decapitated
Trunk blood is immediately collected in EDTA-treated tube.
Remove cells from plasmaion
Centrifuge collected blood in EDTA-treated tube at 2,000 x g for 10 minutes at 4C
Following centrifugation, transfer liquid component into clean polypropylene tube
Aliquot .5ml of isolated plasma
Plasma can be stored at this time at -20C
Denature plasma for immunoblotting
Dilute plasma 1:1 in 1X PBS
Add 4X denaturing buffer to diluted sample
Boil sample for 30 minutes
a. *Due to the high protein content of plasma, increasing denaturing boil time aids in protein separation
Samples are ready for protein loading
a. For best success, leave empty lane (1X denaturing buffer only) between plasma samples