I want to get some helps from all of you...
I have done a lot of SDS-PAGE with "strange" migration of protein near to front dye.
In a normal condition, the smallest protein marker should travel a bit slower than the protein samples migration indicator (bromophenol blue), i get a nice result last time (as shown in normal.png) but after i have prepared a new solution of acrylamide, the condition coming weird.
1. Smallest MW of my protein marker is 10 kDa, it will travel along with the samples indicator in a line, causing the bands near to the marker is distorted and cant get a good result. Somemore, during the destasining process, 1cm broad line apart from the "indicator blue line" is easily washed off and become transparent, so it seems abnormal.
2. The other condition is the smallest MW protein will travel slower than protein marker bigger than it and causing 2 markers were combined between each other.
Is it something happen in the polymerisation process?
Or acylamide contamination?
I tried to change the solution again and again and it doesn't helps.
So, now i m frustrating doing SDS-PAGE and hope to get your advises before i proceed it blindly.