Immunofluorescence Protocol for Protein Localization

Chicken pAb anti-GFP, mAb anti-GAPDH, and mAb anti-N-terminal ERC1a (ELKS-30) against residues 21–40 of mouse/human ERC1a (Abcam, Cambridge, UK); rabbit pAb anti-GFP (Thermo Fisher Scientific, Paisley, UK); rabbit pAbs anti-FLAG and anti-LL5β, mAbs anti-α-actinin, anti-tubulin, and anti-talin (Sigma-Aldrich); mAb anti-vinculin (Upstate, Lake Placid, NY, USA); rabbit pAb against the C-terminus of liprin-α1 (amino acids 818–1202)^{36 (link)} pAb rabbit anti-liprin-α1 (Protein Tech, Chicago, IL, USA); mAbs anti-paxillin and anti-GIT proteins (BD Bioscience, San Jose, CA, USA); mAb anti-LL5 (cl. IH12) from J. Sanes^{39 (link)}; mAb anti Src cl. 327 from S. Courtneidge^{40 (link)}. Alexa-Fluor–conjugated secondary antibodies (Thermo Fisher Scientific, Paisley, UK); horseradish peroxidase (HRP)-conjugated secondary antibodies (Amersham Bioscience, Little Chalfont, UK). Lipophilic dye Nile Red (Sigma Aldrich, St. Louis, MO, USA).