Chapter title |
Monitoring Wnt Protein Acylation Using an In Vitro Cyclo-Addition Reaction.
|
---|---|
Chapter number | 2 |
Book title |
Wnt Signaling
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-6393-5_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6391-1, 978-1-4939-6393-5
|
Authors |
Rubina Tuladhar, Nageswari Yarravarapu, Lawrence Lum |
Editors |
Quinn Barrett, Lawrence Lum |
Abstract |
We describe here a technique for visualizing the lipidation status of Wnt proteins using azide-alkyne cycloaddition chemistry (click chemistry) and SDS-PAGE. This protocol incorporates in vivo labeling of a Wnt-IgG Fc fusion protein using an alkynylated palmitate probe but departs from a traditional approach by incorporating a secondary cycloaddition reaction performed on single-step purified Wnt protein immobilized on protein A resin. This approach mitigates experimental noise by decreasing the contribution of labeling from other palmitoylated proteins and by providing a robust method for normalizing labeling efficiency based on protein abundance. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 2 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 1 | 50% |
Student > Bachelor | 1 | 50% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 1 | 50% |
Agricultural and Biological Sciences | 1 | 50% |