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38 KiB
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{"experiment":{"id":497,"name":"SDS-PAGE for Protein Characterization","description":"This experiment template will help you separate proteins based on their molecular weight. \r\nProteins are separated by electrophoresis through a gel matrix. Smaller proteins migrate faster due to less resistance from the gel matrix. Other influences on the rate of migration through the gel matrix include the structure and charge of the proteins. In SDS-PAGE, the use of sodium dodecyl sulfate (SDS, also known as sodium lauryl sulfate) and polyacrylamide gel largely eliminates the influence of the structure and charge, and proteins are separated solely based on polypeptide chain length.\r\nStaining with Coomassie blue dyes is commonly used to stain proteins and visualize them. But if you want to identify specific proteins (using specific antibodies) a better technique to use is a Western blot.","project_id":223,"created_by_id":202,"last_modified_by_id":202,"archived":false,"archived_by_id":null,"archived_on":null,"restored_by_id":null,"restored_on":null,"created_at":"2018-12-24T10:56:55.416Z","updated_at":"2019-01-28T15:15:24.331Z","workflowimg_file_name":"wimg20181224-1-1hyikrd.png","workflowimg_content_type":"image/png","workflowimg_file_size":2244,"workflowimg_updated_at":"2019-01-28T15:15:24.331Z","uuid":"6d9f073d-dade-4bc3-a27b-2a69b15b8680"},"my_modules":[{"my_module":{"id":2388,"name":"Western Blot","due_date":null,"description":null,"x":99,"y":0,"my_module_group_id":1195,"created_at":"2018-12-24T13:08:01.834Z","updated_at":"2018-12-24T13:43:31.275Z","archived":false,"archived_on":null,"created_by_id":202,"last_modified_by_id":202,"archived_by_id":null,"restored_by_id":null,"restored_on":null,"nr_of_assigned_samples":0,"workflow_order":3,"experiment_id":497,"state":"uncompleted","completed_on":null,"electronic_signature_status":"unlocked","electronic_signature_status_locked_at":null},"outputs":[],"my_module_tags":[],"task_comments":[],"my_module_repository_rows":[],"user_my_modules":[],"protocols":[{"protocol":{"id":3905,"name":null,"authors":null,"description":null,"added_by_id":null,"my_module_id":2388,"team_id":1,"protocol_type":"unlinked","parent_id":null,"parent_updated_at":null,"archived_by_id":null,"archived_on":null,"restored_by_id":null,"restored_on":null,"created_at":"2018-12-24T13:08:01.843Z","updated_at":"2019-01-31T09:18:18.471Z","published_on":null,"nr_of_linked_children":0},"protocol_protocol_keywords":[],"steps":[{"step":{"id":5262,"name":"Transfer of proteins to a membrane","description":"\u003c!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\"\u003e\n\u003chtml\u003e\u003cbody\u003e\n\u003cp\u003eSoak a piece of \u003cstrong\u003ePVDF membrane\u003c/strong\u003e slightly larger than the gel in methanol for \u003cstrong\u003e1 minute\u003c/strong\u003e (pre-wetting), and then equilibrate in transfer buffer. Also, equilibrate two pieces of filter paper slightly larger than the PVDF membrane in transfer buffer. There is no need for pre-wetting a nitrocellulose membrane. Skip using methanol, and equilibrate the membrane in transfer buffer.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eTransfer buffer:\u003c/strong\u003e\u003c/p\u003e\n\u003cul\u003e\n\u003cli\u003e\n\u003cstrong\u003e25 mM\u003c/strong\u003e Tris-HCl\u003c/li\u003e\n\u003cli\u003e\n\u003cstrong\u003e192 mM\u003c/strong\u003e glycine\u003c/li\u003e\n\u003cli\u003e\n\u003cstrong\u003e20%\u003c/strong\u003e MeOH\u003c/li\u003e\n\u003c/ul\u003e\n\u003cbr\u003eEquilibrate the gel in transfer buffer after electrophoresis. \u003cbr\u003e\u003cbr\u003ePlace a piece of equilibrated filter paper on the anode plate, and place the membrane, gel, and another piece of filter paper without any air bubbles. All of these should be sandwiched between two sponges.\u003c/body\u003e\u003c/html\u003e","position":0,"completed":false,"completed_on":null,"user_id":202,"created_at":"2018-12-24T13:17:10.122Z","updated_at":"2019-01-31T09:18:18.436Z","last_modified_by_id":202,"protocol_id":3905},"checklists":[],"step_c
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