combining the results from different gels in western blot - (Jun/04/2008 )

Hi everyone,
I use western blot to compare the levels of specific proteins under two different conditions. I use alfa-tubulin to normalize the results (using the labworks software). I am wondering how I can combine the data that I get from one set of experiments to other? Suppose that I have 5 different gels and therefore 5 different membranes. How can I combine the results for a specific protein from all these membranes?
Thanks
Sayeh

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So long as you have tubulin normalization, you should be okay. Given, of course, that these are same cells and treated with similar conditions. You can cut and paste western blot lanes (keeping slight spaces in between) and/or make a graph with quantification numbers as separate bars or one bar with error bars.

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Thanks for the reply. What if a different exposure time or different antibody concentration is used for different membranes (resulting in different IOD values for different gels)? How would you correct for that? How would you be able to combine all the data and show them in just one bar?

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there is no problem for your normalization if you Westernblot system is calibrated and you know the linear phase of immunosignal; autoluminographic films reach relatively quick saturation; deeper linearity is achieved by densitometer

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