My objective is to extract and purify certain antigen specific chicken egg yolk antibodies. I have found through literatures that, some of the researchers used linear gradient SDS gel to detect purified egg yolk antibodies, some of them have used normal 10 percent or 12 percent SDS gel. Which should I go for?
If you are using reducing conditions, there will be bands at 65-70 kDa and 20-30 kDa. The heavy chain band will run within almost any percentage gel. Whether the light chain band stays within the gel or runs at the front will depend on the gel system you are using. If you are buying precast gels, check the manufacturer's information for where bands of those sizes run on their gels. If you are planning to cast the gel yourself, it is easier to make single-percentage gels than gradient gels, and a 12% gel is a better choice than a 10% gel for this purpose, considering the low molecular weight of the light chain.
If you are not planning to reduce the protein, it will run at about 180 kDa, which means it will be near the top of the gel. A low percentage gel is better for that purpose, either the 4-12% or the 10% would be better than the 12%.
Dear Dutta,
You can go through 10 or 12% SDS-Page gel. If you go with 12% it will be better. Once you run the gel you will get HC heavy chains and LC-light chains. 12% SDS-gel resolve both HC and LC chains. If you have a diluted sample You can go with discontinuing gel [stacking (4-5%) and resolving (12%) gel]
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