How could you demonstrate that MIG1 moved within the cell in response to glucose?
experiment
Answers:
Here's the VERY quick answer. Given the nature of the question I guess you are at Uni and so can fill in the gaps :-)
1. Culture cells with a functional MIG1 pathway
2. Starve the cells of glucose (obviously not totally, and supplement with fructose)
3. Harvest some cells and do something like immunohistochemistry to identify the location of the MIG1 within the cells
4. Add Glucose to the cells
5. Harvest at 1 hr, 2hrs, 4hrs, 8hrs, 12hrs and maybe even later and repeat the immuno
--> hopefully you'll see the MIG1 migrate from one cellular location/compartment to another
If this is for an essay don't forget to explain your controls (most students do and without suitable controls your experiment proves nothing!) For example, you should include a cell line which expresses MIG1 but doesn't have a functional MIG1 pathway.
Good luck and comeback if you have any more specific questions.
YOU HAVE LOST ME HERE !!!! :/
The alternative is to create a GFP-tagged clone of MIG1 and use real-time confocal laser microscopy. You can stimulate your cells under the microscope and then video the real-time movement of the MIG1-GFP in response.
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Answers:
Here's the VERY quick answer. Given the nature of the question I guess you are at Uni and so can fill in the gaps :-)
1. Culture cells with a functional MIG1 pathway
2. Starve the cells of glucose (obviously not totally, and supplement with fructose)
3. Harvest some cells and do something like immunohistochemistry to identify the location of the MIG1 within the cells
4. Add Glucose to the cells
5. Harvest at 1 hr, 2hrs, 4hrs, 8hrs, 12hrs and maybe even later and repeat the immuno
--> hopefully you'll see the MIG1 migrate from one cellular location/compartment to another
If this is for an essay don't forget to explain your controls (most students do and without suitable controls your experiment proves nothing!) For example, you should include a cell line which expresses MIG1 but doesn't have a functional MIG1 pathway.
Good luck and comeback if you have any more specific questions.
YOU HAVE LOST ME HERE !!!! :/
The alternative is to create a GFP-tagged clone of MIG1 and use real-time confocal laser microscopy. You can stimulate your cells under the microscope and then video the real-time movement of the MIG1-GFP in response.
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