{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Cookis T"],"funding":["NIGMS NIH HHS"],"pubmed_abstract":["Streptavidin affinity grids provide strategies to overcome many commonly encountered cryo-electron microscopy (cryo-EM) sample preparation challenges, including sample denaturation and preferential orientations that can occur due to the air-water interface. Streptavidin affinity grids, however, are currently utilized by few cryo-EM labs because they are not commercially available and require a careful fabrication process. Two-dimensional streptavidin crystals are grown onto a biotinylated lipid monolayer that is applied directly to standard holey-carbon cryo-EM grids. The high-affinity interaction between streptavidin and biotin allows for the subsequent binding of biotinylated samples that are protected from the air-water interface during cryo-EM sample preparation. Additionally, these grids provide a strategy for concentrating samples available in limited quantities and purifying protein complexes of interest directly on the grids. Here, a step-by-step, optimized protocol is provided for the robust fabrication of streptavidin affinity grids for use in cryo-EM and negative-stain experiments. Additionally, a trouble-shooting guide is included for commonly experienced challenges to make the use of streptavidin affinity grids more accessible to the larger cryo-EM community."],"journal":["Journal of visualized experiments : JoVE"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11293040"],"repository":["biostudies-literature"],"pubmed_title":["Streptavidin-Affinity Grid Fabrication for Cryo-Electron Microscopy Sample Preparation."],"pmcid":["PMC11293040"],"funding_grant_id":["R35 GM127018"],"pubmed_authors":["Nogales E","Han BG","Cookis T","Glaeser R","Poepsel S","Herbst DA","Sauer P"],"additional_accession":[]},"is_claimable":false,"name":"Streptavidin-Affinity Grid Fabrication for Cryo-Electron Microscopy Sample Preparation.","description":"Streptavidin affinity grids provide strategies to overcome many commonly encountered cryo-electron microscopy (cryo-EM) sample preparation challenges, including sample denaturation and preferential orientations that can occur due to the air-water interface. Streptavidin affinity grids, however, are currently utilized by few cryo-EM labs because they are not commercially available and require a careful fabrication process. Two-dimensional streptavidin crystals are grown onto a biotinylated lipid monolayer that is applied directly to standard holey-carbon cryo-EM grids. The high-affinity interaction between streptavidin and biotin allows for the subsequent binding of biotinylated samples that are protected from the air-water interface during cryo-EM sample preparation. Additionally, these grids provide a strategy for concentrating samples available in limited quantities and purifying protein complexes of interest directly on the grids. Here, a step-by-step, optimized protocol is provided for the robust fabrication of streptavidin affinity grids for use in cryo-EM and negative-stain experiments. Additionally, a trouble-shooting guide is included for commonly experienced challenges to make the use of streptavidin affinity grids more accessible to the larger cryo-EM community.","dates":{"release":"2023-01-01T00:00:00Z","publication":"2023 Dec","modification":"2025-04-19T20:45:18.223Z","creation":"2025-02-19T02:27:09.114Z"},"accession":"S-EPMC11293040","cross_references":{"pubmed":["38224121"],"doi":["10.3791/66197"]}}