MBIO 3460 Lecture Notes - Lecture 10: Nail Polish, Biotin, Green Fluorescent Protein
10 Jan 2018
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Wash and counterstain nucleus: we counterstain the nucleus to see what relevance the fluorescent protein has in contact with the cell. Growth on slides: best for adgerent cells. Chamber slides (coverslip bottom: appropriate for either cell type, allows you to treat cells and image live cells. Storage: short term storage at 4 degrees (up to 2 weeks, long term storage, seal with nail polish and store at -20 (up to 6 months) Basic protocol for fluorescent staining of paraffin embedded samples. Antigen exposure for tissues: citric acid and a pressure cooker or enzymatic. Blocking agents: bsa, serum (goat, bovine, horse) Storage: short term storage at 4 degrees (up to 2 weeks, long term storage, seal with nail polish and store are -20 (6 months) Controls: positive and negative controls, always use a tissue or cell type known to express your protein of interest, a no primary control is very important.
