Ligand and Carbohydrate Engagement (LACE) Assay and Fluorescence Quantification on Murine Neural Tissue

The interaction between cell surface heparan sulphate and diffusible ligands such as FGFs is of vital importance for downstream signaling, however, there are few techniques that can be used to investigate this binding event. The ligand and carbohydrate engagement (LACE) assay is a powerful tool whic...

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Bibliographic Details
Published inBio-protocol Vol. 11; no. 6; p. e3952
Main Authors Clegg, James M, Pratt, Thomas
Format Journal Article
LanguageEnglish
Published United States Bio-Protocol 20.03.2021
Bio-protocol LLC
Subjects
Biology
Methods
HSPG
FGF
LACE
Ligand
Carbohydrate
Heparan Sulphate
FGFR
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Summary:The interaction between cell surface heparan sulphate and diffusible ligands such as FGFs is of vital importance for downstream signaling, however, there are few techniques that can be used to investigate this binding event. The ligand and carbohydrate engagement (LACE) assay is a powerful tool which can be used to probe the molecular interaction between heparan sulphate and diffusible ligands and can detect changes in binding that may occur following genetic or pharmacological intervention. In this protocol we describe an FGF17:FGFR1 LACE assay performed on embryonic mouse brain tissue. We also describe the method we have used to quantify changes in fluorescent LACE signal in response to altered HS sulphation.
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ISSN:2331-8325
2331-8325
DOI:10.21769/BioProtoc.3952