BIO 110 Lecture Notes - Lecture 21: Distal Convoluted Tubule, Fluorescent Tag, Primary And Secondary Antibodies

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You have purified a protein from the ribosomes of a mouse. You want to show whether the protein is responsible for some ribosomes being on the rough endoplasmic reticulum while other ribosomes are free in the cytoplasm. To show if the protein is present on both types of ribosomes or only on one type, inject your isolated protein in a rabbit, isolate the antibody and label with a fluorescent tag. Take mouse tissue, homogenize and conduct cell fractionation: you will have pellets of nuclei, mitochondria, rer and other membranes, free ribosomes, and free cytoplasm. Solubilize the protein in the pellets, run the protein on gel electrophoresis, transfer protein to a membrane support and expose to the fluorescent antibody (i. e. do a western blot). If you see a fluorescent band in both the rer and free ribosome lanes, then the protein is present in both types of ribosomes.

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