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Problem set 3 questions

Problem set 3 questions - CELL BIOLOGY PROBLEM SET 3...

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CELL BIOLOGY PROBLEM SET 3 Answers 1. Define and/or describe the following: a. 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. b. Density gradient centrifugation – Density gradient centrifugation is a method which scientists use to separate intact organelles from a cell. Each type of organelles in a cell has a specific density and based on that density you can separate a certain type of organelle from the others. c. Protein A immunoprecipitation Protein A immuno precipitation o Technique used to purify proteins and collect proteins using an antibody and Protein A. o Protein A comes from staphaureus o How do you collect cultured proteins? Put into test tube. Put in Protein A, attached to a Bead. Protein binds, spin and collect protein d. Western Blots Western Blot o Combination of Gel electrophoresis and ‘blotting’ o Try to identify a protein using an antibody probe. o Process Run gel, flip over. Transfer onto transfer paper, which is like a sponge, using a power supply. Power supply drives proteins out of gel onto paper. Superficially looks like gel. Use indirect technique to add antibody with a tag. Quantitative; you can see an actual amount. e. SELDI-TOF/MALDI-TOF SELDI-TOF o Surface enhanced Laser Desorption Ionization o Process A Protein chip with design at your discretion Add a protein, add less than 1 microliter. Because you need so little sample, you can use for non invasive analysis of tissue fluids.
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Need a Laser to point on protein and proteins ‘fly’ off of plate.
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