Mar 27, 2025

Public workspaceProcessing Blood Intended for Olink Assay

DOI
dx.doi.org/10.17504/protocols.io.261ger81jl47/v1
  • Adrien Stanley1
  • 1Allen Institute for Neural Dynamics
  • Allen Institute for Neural Dynamics
Abbie Weaver
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DOI: dx.doi.org/10.17504/protocols.io.261ger81jl47/v1
Protocol Citation: Adrien Stanley 2025. Processing Blood Intended for Olink Assay. protocols.io https://dx.doi.org/10.17504/protocols.io.261ger81jl47/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: January 27, 2025
Last Modified: March 27, 2025
Protocol Integer ID: 119654
Keywords: Olink assay, Blood preparation, Plasma
Abstract
This protocol describes blood preparation steps required for analysis of cytokines using an Olink assay. This protocol is applied to samples collected during the protocol 'Temporal Assessment of Immune Response.'
Materials

MaterialManufacturer/SupplierCatalog Number
Blood collected for analysis, on ice.
Ice & Ice bucketVWR 76302-992
CentrifugeEppendorfModel# 5424 R
Low protein binding tubesFisher Scientific23-045-258
Caps for low protein binding tubesFisher Scientific22-038-265
Skirted 96 well plateFisher ScientificAB-0800
20-200ul PipetteRainin17014391
20ul Pipette tipsMettler-Toledo17014960

Equipment
Centrifuge
NAME
Eppendorf
BRAND
5427 R
SKU
https://www.eppendorf.com/us-en/Products/Centrifugation/Microcentrifuges/Centrifuge-5427R-p-PF-1322725
LINK
ReagentSimport™ Scientific Micrewtube™ Microcentrifuge Tubes with Low Adhesion SurfaceThermo Fisher ScientificCatalog #23-045-258

ReagentSimport™ Scientific Nonsterile Screw CapsThermo Fisher ScientificCatalog #22-038-265

ReagentThermo Scientific™ PCR Plate, 96-well, low profile, skirtedThermo Fisher ScientificCatalog #AB-0800

ReagentPipet-Lite LTS Pipette L-200XLS+RaininCatalog #17014391

ReagentPipette Tips TR LTS 20µL S 960A/10Mettler ToledoCatalog #17014960


Safety warnings
  • Wear appropriate PPE required by EHS. Proper PPE includes lab coat, eye protection, and disposable gloves.
Ethics statement
This protocol has been approved by the Allen Institute Animal Care and Use Committee (IACUC).
PHS Assurance: D16-00781
AAALAC: Unit 1854
Before start
  • Before starting, set centrifuge temperature to 4°C and ensure lid is closed.
  • Collected blood should be in K3 EDTA coated tubes stored on ice.
  • Blood processing should begin within 30 minutes of blood collection.
Protocol
Temporal Assessment of Immune Response
NAME

Temporal Assessment of Immune Response

CREATED BY
Abbie Weaver

Steps
Steps
40m
40m
Within Duration00:30:00 of blood collection, bring samples to centrifuge and spin at Centrifigation2000 x g for Duration00:10:00 at Temperature4 °C .

40m
Centrifigation
Once centrifugation is complete, place samples TemperatureOn ice or in a rack TemperatureOn ice while trying not to disturb the blood cell pellet.
Remove the top translucent plasma layer using a 20-200ul pipette and pipette tip, then transfer to a non-protein binding tube or a well of a 96 well plate intended for shipment to Olink.

Note
  • Avoid drawing up red blood cells (red pellet at the bottom of the tube), if you do repeat steps 1-2.
  • Olink recommends loading at least 10-20 uL of plasma per well.

Store samples in Temperature-80 °C freezer.
Note
Samples remain stable in these conditions for at least 12 months.



Temperature