Squished lanes in SDS Page
Posted 14 September 2010 - 03:00 AM
got the following prob:
Run 9 samples (all processed the same way) in SDS Page under following conditions:
Nupage Bis-Tris 4-12% precast gels and MOPS buffer from Invitrogen
The samples are cells washed once in PBS, lysed in RIPA + PIs, added Laemmli (reducing)
cooked 5min 99deg, chilled on ice and loaded onto gel
And what I get is 7 of 9 samples running very strangely on the gel (have a look at attached pic).
First thought samples were degraded but....
Only thing I didn't do is spin after cooking the samples
Or too high/low salt concentration?
Posted 17 September 2010 - 08:35 AM
you can reduce salt in a small aliquot for sds-page by drop dialysis.
Edited by mdfenko, 17 September 2010 - 08:37 AM.
genius does what it must
i do what i get paid to do
Posted 17 September 2010 - 02:43 PM
I always had an alternate hypothesis....
Posted 18 September 2010 - 08:10 PM
The OP is using pre-cast gels and a pre-made buffer system from Invitrogen, it shouldn't be either of the problems you mention.
to me it doesnt look lik the prob in the sample or its prep, but the prob is in gel casting or running condn.....