rkay447, on 16 November 2010 - 08:32 AM, said:
Although this is a suggested step, I never did it and never had any issues with my western blots. Perhaps if you do your blot and have background issues, it would be a good idea for the next time. If you do choose to dry your membranes, you should rehydrate before blocking. The instructions with my PVDF (Millipore, Immobilon-FL) says to re-wet the dry blot in 100% methanol for 15 seconds before placing in blocking. From what I've read, the purpose of drying the membrane is so that the proteins are more adherent to the membrane. This might be especially important if you know you are going to need to strip the membrane to get all your desired blots but again, I've never had any problems with membranes that I didn't dry, even after stripping.
Thank u, i have millipore immobilon P and the instructions say that drying is neccesary before any transillumination or rapid immunodetection procedures..So i don't think i have to dry it before blocking correct? I usually dry it approximately 10 min at 37 C and then re-wet with PBST and i don't get noise. But i don't know if that's correct.