While characterising purified antibody on SDS PAGE I ended up with only one band of approximately 14kDa, similar to the J chain, though no heavy or light chains. Does anyone know if you have the J chain present without the main heavy and light ones? More specifically if the purification method used was actually meant to bind to the light chain?
I think you are not providing enough information to answer your questions. It would be useful to know, what is the production system you use to produce your IgMs and what is the method used for purification?
It is possible to detect J chains on SDS- PAGE, but if what you loaded were indeed IgMs, you should be able to see also bands for HC and LC (and those should be much thicker than the band for JC). I would absolutely go for Western blot though, because with the size of ~14 kDa, it is well possible that you only have some kind of contamination not an IgM. It sounds to me as if the purification failed and you didn't elute IgM at all.
I assume you are producing secreted pentameric IgMs? Then alternatively, you could try non-reducing gradient 4-12% PAGE gel to run your samples. There you would be able to see pentamers/hexamers and all the other smaller fractions and you would see pretty clearly if what you purified are IgMs.
I would refer you to a protocol which I have used you with camels, elephants and ostrich it proved to be good. Check my list of publications, Good luck.!!!
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