Hi all,
I am using Oligos to kick down a protein and look on the same time what happens to others as well. I have 6 pairs with normal and corresponding treated cells. I do the transfection with Lipofectin (which I know it is kind of toxic for the cells).
I run yesterday a gel with my samples and loaded the treated ones next to the control ones. The funny thing is that the treated ones as they run in the gel, it seems as they squeeze the normal lanes to the edge, so that at the end the normal lanes are thin and the treated ones are wider. This looks like a yes/no pattern with every second lane to be wider.
Does anybody have an idea what the problem is????
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3 replies to this topic
#2
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an elder
Posted 05 October 2009 - 07:24 AM
it is probably caused by differences in the buffers in which the samples are suspended (prior to addition of laemmli sample buffer). detergents (and other surfactants) and salts can cause strange behavior of the sample if in high enough concentration.
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genius does what it must
i do what i get paid to do
#3
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Enthusiast
Posted 05 October 2009 - 08:05 AM
If you are talking about running a protein gel, one of the problems could be that the treated samples are more concentrated than your normal samples or a component in those samples is more concentrated, causing those lanes to bulge out.
Are you loaded the same amount of protein per lane?
AussieUSA.
Are you loaded the same amount of protein per lane?
AussieUSA.
#4
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Enthusiast
Posted 06 October 2009 - 10:20 AM
[quote name='AussieUSA' date='Oct 5 2009, 09:05 AM' post='38804']
If you are talking about running a protein gel, one of the problems could be that the treated samples are more concentrated than your normal samples or a component in those samples is more concentrated, causing those lanes to bulge out.
Are you loaded the same amount of protein per lane?
AussieUSA.
Hi all and thx for the feedback,
I use exactly the same lysis buffer for every step. The only difference is that my cells are treated and control ones. I think that my problem is sample related but I havent figured out yet what exactly.
If you are talking about running a protein gel, one of the problems could be that the treated samples are more concentrated than your normal samples or a component in those samples is more concentrated, causing those lanes to bulge out.
Are you loaded the same amount of protein per lane?
AussieUSA.
Hi all and thx for the feedback,
I use exactly the same lysis buffer for every step. The only difference is that my cells are treated and control ones. I think that my problem is sample related but I havent figured out yet what exactly.
