2-D Gel Background

[admin]

Hello I got a question by email:

Dear all....

I'm facing a problem with my 2-D Gel , Actually most of the time I have the
background at the acidic side of the gel , and Idon't know what the
resons for that while the other side of the gel it's clear.So, any one know what
the reasons ?
thank u for your help

[danfive]

For Background--not smearing, dropping out, smudging--washing your gel before staining helps a great deal. You can rinse and wash for 30 minutes to 1 hour--I suggest rinsing 4 to 5 times in 4X volume--then washing on a shaker gently for 30-60 minutes--then using a minimal amount of time for staining. Before visualization rinse again. Also use new--not reused staining solution. Some people rinse and then fix gel with a TCA solution--not good for MS downstreaming though---I find this to be overkill.