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Question: question 10 1 pts determining the number of transmembrane domains...

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Question 10 1 pts Determining the number of transmembrane domains in a protein You have recently identified a novel protein of the plasma membrane in yeast cells. This protein has the following hydrophobicity plot. 2 -1 -2 0 20 40 60 80 100 120 140 160 You Pl suggests to use immunofluorescence microscopy to determine the orientation of this protein in the membrane. He gives you the following 4 antibodies: 1. antibody to the N-terminus, raised in rabbit 2.goat anti-rabbit secondary antibody with a red fluorophore 3. antibody to the C-terminus, raised in mouse 4.goat anti-mouse secondary antibody with a green fluorophore If you use detergent, you see a yellow signal at the plasma membrane, which means colocalization of the red and the green signal. If you dont use detergent, you only see a red signal at the plasma membrane. Which conclusion can you draw about the organization of this protein in the membrane? Note: The preparation of cells for immunofluorescence analysis requires a chemical fixation step (often formaldehyde or methanol), followed by permeabilization of membranes with detergent. This permeabilization step helps antibodies get access to their antigen inside of the cell. The protein has two transmembrane domains, and N-and C-termini are both in the exoplasmic space (outside of the cell). O The protein has two transmembrane domains, and N-and С-termini are both in the cytosol The protein has three transmembrane domains, and N- terminus is in the cytosol. The protein has three transmembrane domains, and N-terminus is in the exoplasmic space (outside of the cell) Immunofluorescence cannot be used to determine the orientation of a protein in the membrane.

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