slow electrophoresis, somebody can help me? - troubles in electrophoresis (Oct/06/2008 )
Hello everybody:
We have a problem with western blot...I don't know why, but, the last times our electrophoresis was very very slow... (4 hrs!!!!) in a 10% SDS-PAGE gel. I tried to identify the problem but it's difficult, i tested the electrical supply and electrodes and all seem to be ok.
Has somebody some idea or suggestions???
Thanks in advance.
Jose.
was your running buffer freshly prepared? And prepared correct? If there are not enough salts in it your gel will not run at all. Did you fill the buffer also high enough in the inner chamber between the gels so that the top of the gel is in contact with the buffer? Did this happen before or was it only one time? Perhaps, next time everything will go on as usual. Otherwise keep an eye on every detail.
We have a problem with western blot...I don't know why, but, the last times our electrophoresis was very very slow... (4 hrs!!!!) in a 10% SDS-PAGE gel. I tried to identify the problem but it's difficult, i tested the electrical supply and electrodes and all seem to be ok.
Has somebody some idea or suggestions???
Thanks in advance.
Jose.
some possibilities:
- apparatus out of order (but you checked it)
- too much salt in samples
- too much salt in sample or running buffer (10x stock instead of 1x used?)
- leakage in the system, short circuit between cathode and anode buffer
Thanks a lot biomaus, today I'll prepare new running buffer and...I hope that the problem will be resolved.
Jose.
We have a problem with western blot...I don't know why, but, the last times our electrophoresis was very very slow... (4 hrs!!!!) in a 10% SDS-PAGE gel. I tried to identify the problem but it's difficult, i tested the electrical supply and electrodes and all seem to be ok.
Has somebody some idea or suggestions???
Thanks in advance.
Jose.
I have had the same problem, and the problem vas old Glycine (not special for electrophoresis) in buffer.