horizontal streaking in 2-D gel? - for help (Jul/19/2007 )

why are the horizontal streaking present in the alkaline part of the 2-D gel (3-10)?

is the possible cause underfocusing or overfocusing? or are there any other reasons?

the left in the figure is the alkaline part.

once can i do a good job in 2-D gel with the same methods for the same sample, but now???

please advise !

xiaofei

xfzhang04@genetics.ac.cn

the picture is just the alkaline part the the whole 2-d gel. there is an interesting phenomena : the right part of spots are good, but the left part are streaking. why? can i get a good figure by increasing the time of focusing?


looks like uncomplete focussing of the 1st D; increase time of focussing
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there are three factors i can think of:

incomplete focussing, increase time (as the bearer said);

overloaded sample, decrease amount of sample loaded;

and

aggregated and denatured proteins, clarify sample immediately prior to application

or any combination of these three.