Immunofluorescence of GFAP and TNF-α in the Mouse Hypothalamus.
Cryosections
Glial Fibrillary Acidic Protein (GFAP)
Hypothalamus
Immunofluorescence
Neuropeptides
TNF-α
Tumor necrosis factor alpha
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
05 Jul 2021
05 Jul 2021
Historique:
received:
26
01
2021
revised:
15
04
2021
accepted:
24
04
2021
entrez:
30
7
2021
pubmed:
31
7
2021
medline:
31
7
2021
Statut:
epublish
Résumé
Immunofluorescence is a reliable method for identifying specific proteins in neuronal and glial cell populations of the hypothalamus. Several immunofluorescence protocols are available to detect protein markers and neuropeptides in the hypothalamus; however, published methods may vary in subtle details that can potentially impact the final outcome of the procedure. Here, we provide a detailed protocol suitable for thin cryostat sections, which has been successful for specific antibodies directed against key markers of hypothalamic neurons and glial cells. We include every detail concerning brain tissue collection, processing, sectioning, and labeling with optimal dilutions of antibodies with the aim of reducing non-specific background. Our background-optimized immunostaining protocol has been routinely used in the lab and allows efficient detection of specific neuropeptides, glial cells, and markers of inflammation and endoplasmic reticulum stress in the hypothalamus.
Identifiants
pubmed: 34327275
doi: 10.21769/BioProtoc.4078
pii: e4078
pmc: PMC8292123
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e4078Informations de copyright
Copyright © 2021 The Authors; exclusive licensee Bio-protocol LLC.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare no competing interests.
Références
J Vis Exp. 2012 Jul 30;(65):
pubmed: 22871843
Nat Methods. 2012 Jul;9(7):671-5
pubmed: 22930834
Int J Obes (Lond). 2017 Jan;41(1):149-158
pubmed: 27773938
Bio Protoc. 2018 Jan 20;8(2):
pubmed: 29552595