title: Common fixation-permeabilization methods cause artifactual localization of a type II transmembrane protein.
abstract: We found that a localization artifact can arise from common immunofluorescence methods. Specifically, cell fixation and permeabilization can cause mislocalization of a type II membrane-bound protein, ER mannosidase I, from its native localization in vesicles to the Golgi complex. Live cell microscopy and interestingly also mild cell fixation with paraformaldehyde without membrane permeabilization do not present this artifact.
authors: Department of Cell Research and Immunology, George Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors: Ron
authors: R
authors: Benyair
authors: Department of Cell Research and Immunology, George Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel gerardo@post.tau.ac.il.
authors: Gerardo Z
authors: GZ
authors: Lederkremer
mesh_terms: D016475:3T3 Cells; D000818:Animals; D016477:Artifacts; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000648:ultrastructure*; D005456:Fluorescent Dyes / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D006056:Golgi Apparatus / Q000502:physiology*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008361:Mannosidases / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D011108:Polymers / Q000737:chemistry; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D008565:Membrane Proteins; D011108:Polymers; D005557:Formaldehyde; D008361:Mannosidases; C046090:mannosyl-oligosaccharide 1,2-alpha-mannosidase; C003043:paraform
keywords: Golgi; fixation; mannosidase; permeabilization; transmembrane protein; vesicles