I found that Leptin can form dimers and trimers. I performed my western blot in denaturant conditions with ß-mercaptoethanol and heated at 95ºC, however, it still appearing 2 strips up and down of 31 kDa when leptin MW is 16 kDa. I already used 2 antibodies sc-842 and AB16227. I hope someone can help me.
It looks strange. How much was the concentration of Beta-mercaptoethanol? Did your sample buffer contain SDS? You need to use SDS for complete denaturation of your protein into its monomeric form.
The other possibility might be due to non-specific binding of your antibodies with some host proteins present as impurities in your samples. Are those antibodies sera? or Are they monoclonal antibodies?
Hi, I already checked the protocols and concentration of ß-mescapto and SDS are ok. I run my samples for 2 proteins (adiponectin and leptin) and adiponectin was well denatured. Both are rabbit polyclonal antibodies
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