Dear Researchers,
I was preparing to run 2nd dimension (SDS PAGE) after finishing 1st dimension (isoelectric focusing). After finishing everything perfect, i was sealing top of the IPG strip with 0.5% agarose solution. Unfortunately the solution got solidified while i was pouring. So lot of air bubbles got trapped in between the strip and agarose solution and between the strip and separating gel. As I didn't want to take risk of removing the strip out of the gel i started running the gel as such. I am very much worried whether the bubbles will interfere with protein separation? Experts please help me in this regard.
Hello. I think you must remove the strip because bubbles can cause loss of conductivity being air between your agarose and polyacrylamide gel and therefore make mistakes such as proteins that enter and migrate poorly in the gel.
Hello. I think you must remove the strip because bubbles can cause loss of conductivity being air between your agarose and polyacrylamide gel and therefore make mistakes such as proteins that enter and migrate poorly in the gel.
bubbles will affect the separation. You must have removed the strip and sealed once again.
The separation pattern will distort as the proteins are dragged around the bubbles.
On every single bubbles will appear a huge gap in the 2D pattern...
Sad...but...repeat
yes bubbles do affect protein separation, especially if they are present exactly in the way of your proteins running. Cheers
Even micro-air bubbles generate lots of impedance for current flow, since they are nonconductor of electricity resulting in uneven movement of proteins in the gel. You need to repeat it a fresh!
Yes bubbles affect the separation in 2D-gel and PAGE as well. Separation pattern of protein will distort, you must need to be careful during experiment.
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