feb 10 notes - Anatasia Gangadin Cell Biology Finish...

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Anatasia Gangadin Page 1 10/6/2009 Cell Biology Finish Microscopy Proteins Techniques Thursday FRET Fluorescence Resonance Energy Transfer o Technique that reports back if two molecules are binding to each other (1-10 nanometers) o Relates to Green fluorescent protein. o Process We have a protein. Can wither be a protein X or a drug which was synthesized and we want to know if it binds. We have coupled Blue Fluorescent protein. If you excite BFP with Violet light, light comes out blue. If one had GFP and is excited with blue light then green light is produced. If both were in a cell, and you only see blue then there is no binding. If you see green, there is binding. If bound, in terms of drug or protein binding, it is within 1-10 Nanometers. Other Techniques Autoradiography o Use of radioactive tags to track a cellular process. o Examples I-131 (Radioactive Iodine) Labels the Thyroid gland. Thyroxin is produced, which produces Iodine. H-3 Thymadine DNA Base used to label diving cells because it labels cells going through the S phase of cell life. o Group of cells in culture. Add Tridiated Thymadine. Only labels a cell going through DNA synthesis. Overlay a film emulsion, and once it is layered, put into a dark box. Decay particles expose the film. Cells are shown and a dark one signals DNA synthesis. H-3 Leucine or S-35 Methionine (amino acids) Looking at Protein Synthesis Trydiated Leucine is used as ‘pulse chase’. Radioactive probe is added then washed out because it is incorporated into cell, and then you ‘chase’ it with a non radioactive probe. Takes 30 minutes. After the chase, path can be traced. In situ hybridization
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This note was uploaded on 04/13/2009 for the course BIOL 311 taught by Professor Vanbuskirk during the Spring '08 term at Binghamton University.

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feb 10 notes - Anatasia Gangadin Cell Biology Finish...

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