SDS PAGE aberrant ladder behaviour - SDS PAGE problem (Sep/02/2005 )
Hi
I ran NuPage 4-12% gel with MOPS buffer and LDS sample buffer. The 4th lane had standard (Biorad dual color) while the 5th to 9th had my IP samples. When I stained the gel with silver stain i noticed an odd thing. The standards appearerd up to the 9th lane. I am pretty sure that while loading i did not leak it. Even if it did, then it should be only on the 5th or 3rd lane. Further I also noticed a tiny 
connection between each of this lane but only at the position of each band of the standard. Is it possilbe for the ladder to migrate sideways? My protein samples have not shown this pattern. Also the lanes 1-3 on the left side of ladder did not show this phenomenon. Please help! Due to this problem I cannot cut out my bands for sequencing>
I believe you are using 12 well NuPAGE gel.
If you are reading from side A, the marker is in 4th lane. 1st, 2nd, 3rd, 4th
If you turn it over, side B, (reflective side), the fourth lane is the 9th lane. 12th (1st), 11th (2nd), 10th (3rd), 9th (4th)
The protein will NOT migrate.
I guess you are just looking at the gels from another side.
If you turn it upside up, the 9th lane will changes back to the 4th lane.
Hope this helps!
I ran NuPage 4-12% gel with MOPS buffer and LDS sample buffer. The 4th lane had standard (Biorad dual color) while the 5th to 9th had my IP samples. When I stained the gel with silver stain i noticed an odd thing. The standards appearerd up to the 9th lane. I am pretty sure that while loading i did not leak it. Even if it did, then it should be only on the 5th or 3rd lane. Further I also noticed a tiny
connection between each of this lane but only at the position of each band of the standard. Is it possilbe for the ladder to migrate sideways? My protein samples have not shown this pattern. Also the lanes 1-3 on the left side of ladder did not show this phenomenon. Please help! Due to this problem I cannot cut out my bands for sequencing>