western with nuclear extract - does high salt/glycerol affect running (Feb/23/2007 )
Hi,
I've been preparing nuclear extract with 420mM salt and 25% glycerol.
When I run this on a gel next to whole cell extract it often runs
as a diffuse band or with some smearing.
However, the whole cell extract or in vitro translated products
look great.
Has anyone run into this type of problem before?
-mikew-
QUOTE (mikew @ Feb 23 2007, 08:23 PM)
Hi,
I've been preparing nuclear extract with 420mM salt and 25% glycerol.
When I run this on a gel next to whole cell extract it often runs
as a diffuse band or with some smearing.
However, the whole cell extract or in vitro translated products
look great.
Has anyone run into this type of problem before?
I've been preparing nuclear extract with 420mM salt and 25% glycerol.
When I run this on a gel next to whole cell extract it often runs
as a diffuse band or with some smearing.
However, the whole cell extract or in vitro translated products
look great.
Has anyone run into this type of problem before?
too much salt; precipitate before running SDS-PAGE
-The Bearer-
i prepare nuclear extracts that contain 50% glycerol.
But only 300mM KCl.
-fred_33-