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I figure there's a chance that I can get a FRAP passage so I thought I might as well try to understand it now, than during the DAT (tomorrow). Hopefully by trying to explain it to you guys, I'll understand this a little better, and please feel free to add or correct any of this.
So FRAP- fluorescence recovery after photobleaching.
Purpose: to study mobility of nearby proteins.
How/Experiment: induce/determine/define flourscense level of nearby proteins of interest. Flashed/photobleach a specific target. This will make small area very dark compared to the nearby induced flourscent proteins. After some time, flourscent molecules will diffuse (if mobile) towards and into the bleached/dark area. Eventually nearby flourscent molecules will mix with the bleached out area's visibility/floursence is recovered.
Conculusion: (lol this was not intended to be a lab write up) So diffusional mobility could be studied by monitoring the how fast flourscence returns to the photobleached area. If the fluorscene recovery is fast, nearby proteins are highly mobile ("high % recovery with fast mobility). If the recovery is slow or none, nearby proteins are not mobile ("low % recovery with slow mobility").
Ok, so if the the nearby proteins are anchorered, you're gonna have low % recovery and slow mobility.
If the nearby proteins are large but is not anchored, you will get high % recovery but with slow mobility.
-yada yada yada....you get the pattern.
Wiki it to find the graph image, and hopefully this helps. It helped me atleast =) .
peace.
So FRAP- fluorescence recovery after photobleaching.
Purpose: to study mobility of nearby proteins.
How/Experiment: induce/determine/define flourscense level of nearby proteins of interest. Flashed/photobleach a specific target. This will make small area very dark compared to the nearby induced flourscent proteins. After some time, flourscent molecules will diffuse (if mobile) towards and into the bleached/dark area. Eventually nearby flourscent molecules will mix with the bleached out area's visibility/floursence is recovered.
Conculusion: (lol this was not intended to be a lab write up) So diffusional mobility could be studied by monitoring the how fast flourscence returns to the photobleached area. If the fluorscene recovery is fast, nearby proteins are highly mobile ("high % recovery with fast mobility). If the recovery is slow or none, nearby proteins are not mobile ("low % recovery with slow mobility").
Ok, so if the the nearby proteins are anchorered, you're gonna have low % recovery and slow mobility.
If the nearby proteins are large but is not anchored, you will get high % recovery but with slow mobility.
-yada yada yada....you get the pattern.
Wiki it to find the graph image, and hopefully this helps. It helped me atleast =) .
peace.
