I was measuring the level of protein A production after exposure to Protein B in different 3 conditions e.g. control(no Protein B), wild type and mutant protein B and with treatment with three different drugs therefore there will be 12 different bands including the untreated. I ran untreated and treated control, wild type and mutant samples on the same gel, however it was not possible to run all duplicate or triplicate in the same gel as I have only 15 well gel. Bands were analyzed using ImageJ .Then I did a couple of normalizations. Level of protein A was normalized to respective GAPDH control. The untreated control( no exposure to Protein B and no drug) condition was normalized to 1 and all other experimental conditions were compared to this. How can I do the stats to show the significant difference between the condition as I have only one value per each condition? Thank you in advance.
It will be difficult to justify comparisons between different gels.
Perphaps it may be possible to use a calibrator that would be present in all samples, but you'd have to make sure it's the same intensity in all gels. Even with that, some people may not be convinced. Perhaps opting for a bigger gel may be the best approach.
http://www.bio-rad.com/en-ug/sku/3450051-10-criterion-tbe-polyacrylamide-gel-122-well-45-ul?ID=3450051
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