HI,

Since pro IL-1beta is 33kDa protein, I would recommend using 15% gel concentration. This will also help you detect IL1beta if you want as well. Better would be a gradient gel, but a 15% gel will do the job. Any standard transfer set-up will work in this case. Good luck.

Hi Claudio,

12% - 15% gel concentration is nice for this protein. You may try SDS-PAGE and non-reducing PAGE (without beta-mercaptoethanol, SDS and sample heating). For transfer, it will depends on your buffer and equipament, if you give this information it would be helpfull.

Hi!

Thanks you both for your answer, Leandro Toshio Kochi , Santosh Jadhav

Currently, my conditions are SDS-PAGE in a 15% gel concentration,. For transfer, I dispose of transference buffer consisting in Tris Base/Glycine, methanol (20% in final solution) and distilled water for 1L. As equipament, I have PowerPac Basic Power Supply from BioRad.

I was thinking on time of transfer and set it up to 60 min at 350 mAh, due the molecular weight of pro-IL-1Beta and the cleaved form.

I think the parameters are okay.