Western Blotting: Difference between revisions
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#Turn on heat block to 85 degrees | #Turn on heat block to 85 degrees | ||
#Run SDS-PAGE gel using [[ SDS-PAGE Running Buffer ]] and prepare diluted transfer buffer | #Run SDS-PAGE gel using [[ SDS-PAGE Running Buffer ]] and prepare diluted transfer buffer | ||
## Use a prepared | ## Use a prepared gel (in the 4 degree). Remove it from the packaging, remove the white strip of tape from the bottom back, and gently pull the comb out and rinse with water. Pick a percent gel based on your expected protein size. | ||
##Load into gel tank. Fill with SDS page Running buffer (1X) to the fill line in the front and halfway up the back | ##Load into gel tank. Fill with SDS page Running buffer (1X) to the fill line in the front and halfway up the back. For a video on how to use these gel tanks see https://youtu.be/aedIjTtNEus | ||
##Boil sample at 85 degrees for ~3 min | ##Boil sample at 85 degrees for ~3 min | ||
##Load 3 microliters of protein ladder (purple top) (in the 4 degree), and 10 microliters of each sample into separate wells. | ##Load 3 microliters of protein ladder (purple top) (in the 4 degree), and 10 microliters of each sample into separate wells. | ||