Feb 07, 2024

Congo Red Immunostaining

DOI
https://dx.doi.org/10.17504/protocols.io.4r3l22k1jl1y/v1
  • 1Department of Clinical Neuroscience, Karolinska Institutet, 171 76 Stockholm, Sweden;
  • 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Jacquelyn Haytayan
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DOI: https://dx.doi.org/10.17504/protocols.io.4r3l22k1jl1y/v1
Protocol Citation: daniel.dautan , Per Svenningsson 2024. Congo Red Immunostaining. protocols.io https://dx.doi.org/10.17504/protocols.io.4r3l22k1jl1y/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: February 07, 2024
Last Modified: September 23, 2024
Protocol  Integer ID: 94867
Keywords: ASAPCRN, congo red immunostaining staining for amyloid structure, amyloid structure, congo red immunostaining staining, protein aggregate, protein
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: 020608
Abstract
Staining for amyloid structures in protein aggregates
Wash freshly sectioned tissues 2-3 times with 1X PBS to remove OCT.
Mount all sections onto microscope slides and dry at Room temperature 00:00:00 .
Transfer slides into a 1% CongoRed solution for 00:30:00 .
30m
Place slides into alkaline bath (1% Sodium hydroxide in 50% ethanol) for 00:05:00 .
5m
Transfer slides into 3 sequential baths of water to remove excess staining.
Transfer slides into 0.4% Toluidine blue solution for 00:10:00 .

10m
Wash slides in 3 sequential bath of water to remove excess staining.
Finally, dehydrate slides using sequential baths of:
  1. distilled water (~00:02:00 )
  2. 70% ethanol (2 times ~00:02:00 )
  3. 95% ethanol (2 times ~00:02:00 )
  4. 100% ethanol (2 times ~00:02:00 )
  5. 10% xylene (2 times ~00:05:00 )
13m
Dry sections at room temperature (~00:05:00 ).
5m
Cover with DPX mounting medium.