SDS QUESTIONS: Researchers wanted to study the effect of nicotine on connexin 43
ID: 213004 • Letter: S
Question
SDS QUESTIONS:
Researchers wanted to study the effect of nicotine on connexin 43 levels in endothelial cells of blood vessels. Connexins are proteins that form gap junctions which allows adjacent cells to communicate quickly. The researchers isolated endothelial cells and grew them in vitro. One group of cells was exposed to nicotine for 8 hours while the control cells were not exposed to nicotine. Protein was isolated from each group of cells at the end of the experiment and equal amounts of protein were ran on an SDS- PAGE gel. Connexin 43 will run at 53kDa. I next did a Western blot on these samples using an antibody that would recognize Connexin 43. For your personal info, they then isolated endothelial cells from smokers undergoing heart bypass surgery and got the same results! What does this gel seem to suggest about Connexin 43 regulation in this experimental model? SDS/PAGE Con Exp 53kDaExplanation / Answer
1. Conexins are used to make gap junctions for communication quickly. After addition of nicotine connexin will be increased in the experimental cells. Connexin makes a faster transfer of nicotine to the neighbouring cells.
But control cells no increase in the connexin.
2. Acindin is an enzyme is used to digest the different substrate. Type 1 collagen - 139KDa is digested and moves to end of the gel. May be it has 4 subunits each has 30kDa so it migrated very low. Fibrinogen is 340kDa is digested to make 200kDa and 150kDa with different subunits. So fibrinogen is digested by Acindin.
Laminin is 400KDa the bands also present in 400KDa. so it is not digested by acindin.
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