Mitochondrial Protein Interactions by Cross-link IP
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Corresponding Organization : McGill University
Other organizations : Karolinska Institutet
Variable analysis
- Chemical cross-linking with 1 mM dithiobis-sulfosuccinimidyl propionate (DSP)
- Proteins identified by mass spectrometry
- Mitochondria enriched heavy membrane fractions (200 μg) from control fibroblasts
- Lysis buffer (10 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% n-dodecyl-D-maltoside (DDM), and complete protease inhibitors)
- Non-coated Dynabeads Protein A for pre-clearing
- Antibody binding to Dynabeads Protein A and cross-linking with 20 mM dimethyl pimelimidate (DMP)
- Washing with lysis buffer
- Elution using 0.1 M glycine pH 2.5/0.5% DDM
- Not explicitly mentioned
- Not explicitly mentioned
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