12% SDS-PAGE running strange - (Mar/10/2006 )
Help!
I have a problem, I just started an internship and I'm doing western blottings.
I use 12% SDS-PAGE gells.
Sometimes my gells run very strange, not in a straight line but in curves.
Has anyone experience with this kind of problem? Any ideas of what I'm doing wrong?
I've attached a picture of the failed blot I did today, you can see the strange way my gell ran.
.
P.S.
One time I made 3 gells at the same time, put all the ingredients in one tube. Two (in one tank) ran perfect but the other (in an other tank) ran strange
Could this mean that it's the tank that's the problem? Try a tester using both tanks (with gels made up from the same buffers etc like you did last time) and see if the gel in the tank that produced the strange gel before produces a strange gel again. Could be that the current isn't running correctly in this tank and if so you might need to get it fixed or throw it away and get a new one (but make sure it's definately the tank before throwing it away!)
have you tried running the same samples in both tanks? there could be a component in some of your samples that can cause this problem (eg- high salt, lipids, peg,...).
hi
i think you are leaving a gap between the wells if yes you want to do like that give a support means load a sample buffer 5 ul between the gap wells and also first and last ex: you have 3 samples
samplebuff 1st sample sampbuff 2nd samp samplebuff 3rdsamp sample buff 1 2 3 4 5 6 7
try like this all the best
hi,
in my view, i donot think there is something wrong in salt conc or lipid levels in the sample coz marker also ran absurdly.
since u used same reagents for two gels u r gel composition is nice.
i do agree there is something wrong in gel tank, before this whether u used same running buffer composition for both tanks?
change the apparatuse n try
gud luck
I have a problem, I just started an internship and I'm doing western blottings.
I use 12% SDS-PAGE gells.
Sometimes my gells run very strange, not in a straight line but in curves.
Has anyone experience with this kind of problem? Any ideas of what I'm doing wrong?
I've attached a picture of the failed blot I did today, you can see the strange way my gell ran.
.[attachment=698:attachment]
P.S.
One time I made 3 gells at the same time, put all the ingredients in one tube. Two (in one tank) ran perfect but the other (in an other tank) ran strange
Yes it sounds like one of your tanks has gone a bit wrong, either that or you've not got rid of all the air bubbles from under the gel before starting your run.