12) Use tweezers to put membrane into a small tupperware top and cover with ponso (red stuff). Staining the blot: 11) Dump out the TB, and remove the sandwich.
Western blot sandwich order manual#
10) Run the blot on manual and constant V at 100V for about 1 hour. Then fill the gel box up to the top of the red half of the sleeve with TB. 9) Put the blotting sandwich into the black and red sleeve with the black part of the sandwich facing the black part of the sleeve. Then put the ice pack in next to the sleeve. 8) Put a small stir bar in the gel box underneath the black and red sleeve. 7) Roll a 15ml tube up and down and side to side over the completed sandwich to remove any bubbles.
6) To complete the sandwich, one more piece of paper goes on and then the last pad, again making sure both are thoroughly soaked in TB. When it is completely soaked with TB, move it onto the sandwich next. 5) Use the tweezers to grab the nitrocellulose membrane and dunk it in the dH2O container, make sure the entire membrane is wet, and then move it to the TB. Then put on a pad and one piece of paper (make sure both are completely wet in the TB). 4) To make the blotting sandwich, begin by putting the blotting apparatus in the TB container with the black side down. To remove the other plate, place the gel in the TB container and gently scrap off with a flat scoopula. 3) Remove one glass plate from the gel by twisting a spacer between the plates. Set up blot: 2) Prepare two tupperware containers, one with dH2O and one with 1/2X Transfer Buffer (TB).
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