Air bubbles in PAGE

[tss81exon]

Hi all,
I have problems with air bubbles between gel and the glass plate while casting PAGE. I am not able to figure out the reason for this . Do these air bubbles interfere with protein movement in the PAGE and if it does, can anyone suggest me a way to avoid air bubbles?

Thanks

Silpi

[tshego]

Hi Silpi,

Try adding a 200 uL layer of Absolute 100% Alcohol after loading your gel. This will eliminate the Oxygen from the gel whilst it's polymerizing. After your gel has set, decant the alcohol and rinse with running gel overlay.

tshego

[mai-mb]

I also found saturated butanol works very well ( use as directed above!!).
here is a recipe I use:
Combine 50 ml of n-butanol or isobutanol (or isopropanol) and 50 ml of H2O in a glass bottle and shake. Use the top phase to overlay gels. Store at room temperature indefinitely.

[danto]

air bubbles can affect the movement of protein since when the protein get into contact with the bubble, they revolve around the bubble before continuing to move.position the silage slightly on top of the plate in a slanting position to avoid air from getting inside