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Proteomic workflow and validation. (A) Workflow for the affinity enrichment and analysis of endomembrane proteins from A. thaliana. (B) Immunoblotting of RFP-RABF2b/ARA7 and YFP-RABG3f proteome enrichments to determine organelle contamination. Total protein extracts, from A. thaliana Col-0 or stably expressing RFP-RABF2b/ARA7 or YFP-RABG3f were subjected to immunoaffinity enrichment of RFP or YFP followed by immunoblotting with αRFP, GFP, BIP2, COXII, RbcL, AHA1 as indicated.
