Apr 29, 2026

Blood collection and serum preparation in mice

DOI
https://dx.doi.org/10.17504/protocols.io.kqdg3mnkql25/v1
  • 1Department of pharmacology and physiology, Faculty of Medicine, Université de Montréal;
  • 2Neural Signaling and Circuitry research group (SNC);
  • 3Center for Interdisciplinary Research on the Brain and Learning (CIRCA);
  • 4Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815;
  • 5Department of neuroscience and physiology, Faculty of Medicine, Université de Montréal;
  • 6Institut Courtois d’innovation biomédicale
Amandine Even
Icon indicating open access to content
QR code linking to this content
DOI: https://dx.doi.org/10.17504/protocols.io.kqdg3mnkql25/v1
Protocol Citation: Amandine EVEN, Sriparna Mukherjee, Louis-Éric Trudeau 2026. Blood collection and serum preparation in mice. protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg3mnkql25/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: March 31, 2026
Last Modified: April 30, 2026
Protocol  Integer ID: 314144
Keywords: blood collection from mice, serum preparation in mice, serum preparation, blood collection, multiplex cytokine assay, resulting serum, mice, blood, protocol, preparation
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-000525
Abstract
This protocol describes blood collection from mice and serum preparation. The resulting serum can be used for ELISA or multiplex cytokine assays.
Guidelines
Avoid unnecessary freeze-thaw cycles for serum.
Materials
Drugs and solutions:
- Sodium pentobarbital (7mg/mL in saline)
- PBS 1X (pH~7.5)

Apparatus:
- Refrigered microcentrifuge

Other materials:
- scissors for dissection
- 1.6mL eppendorf tubes
- 1mL syringe with 21G needle
Blood collection
18m
Mice are anesthetized with an intraperitoneal injection of sodium pentobarbital (100 mg/kg ).

15m
Blood is harvested via cardiac puncture with a 1 mL syringe and 21G needle.
This is a terminal procedure and is typically performed immediately before euthanasia or tissue harvest.

2m
Blood is collected in a dry 1.6 mL eppendorf tube.

1m
Serum preparation
37m
Allow blood to clot 00:20:00 at Room temperature .
20m
The sample is centrifuged at 2000 x g, 4°C, 00:15:00 to obtain serum.
15m
Serum is kept at -80 °C until analysis.
For a multiplex cytokine analysis, a 2-fold dilution of serum is prepared with PBS 7.5 prior to freezing.
Label samples with unique animal and group identifiers and keep under conditions compatible with the assay provider’s instructions.
2m