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Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange (ABE)
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Capítulos
- 00:05Título
- 02:08Extraction of Target Protein from Cultured Primary Hippocampal Neurons
- 05:54Acyl-Biotin Exchange: Hydroxylamine (HAM) Cleavage
- 09:17Elution of Biotin-conjugated Target Protein and SDS-PAGE
- 10:46Western Blotting and Detection of Palmitoylation of Target Protein
- 12:16Results: Detecting Palmitoylation of a Neuronal Protein
- 16:07Conclusion
The reversible addition of palmitate to proteins is an important regulator of intracellular protein trafficking. This is of particular interest in neurons where many synaptic proteins are palmitoylated. We utilize a simple biochemical method to detect palmitoylated proteins in cultured neurons, which can be adapted for multiple cell types and tissues.
