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-   -   How to Run a Concentrated Protein Sample for Western Blot? (http://www.molecularstation.com/forum/western-blot-forum/74972-how-run-concentrated-protein-sample-western-blot.html)

eng 01-17-2011 05:59 AM

How to Run a Concentrated Protein Sample for Western Blot?
 
Quote:

Originally Posted by danfive (Post 422718)
Your pic looks fine, I did notice some difference in the lane widths. But that is probably due to running ~17 lanes. That's a lot of samples for this one gel, and that is likely what causing the slight distortions---overall your coomassie stained gel is certainly of acceptable quality.

I wouldn't dilute with water, just a waste, put your serum in sample buffer or Laemmli Buffer.

Hi Danfive and those who may know the answer,
I am doing Western Blot with plasma samples.
I am facing with the problem of too concentrated my plasma sample until 80 ug/ul.
I need to dilute it with Laemmli Buffer?
Using 1 X Laemmli?
Thanks for your kindly attention.

admin 02-18-2011 12:13 AM

Re: How to Run a Concentrated Protein Sample for Western Blot?
 
Hi there,
depending on if you want a more concentrated sample - you can simply lyse your cells in loading or Laemmli Buffer directly. Then load the amount you need.

If you want to run diluted samples, you can simply analyze your samples to find protein concentration - use the Bradford Assay or a similar kit. Then simply dilute the samples accordingly. You can use Laemmli Buffer or loading buffer to do this.


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