Is there is any possibility that my gel will not suffer any damage/alteration in blotting, if I store the gel before western blotting?
It is preferable that the transferring of protein bands exactly after the running of them to prevent diffusion of proteins specially in non purified proteins. Although, you can keep the gel up to 12 hours at 4◦ C.
Hi there,
The risk during storage is diffusion of the bands generating during migration. Therefore bands revealed by WB might be diffuse too. But why not running the transfer step right after SDS/PAGE. Blotted membrane are much more stable than gel (either under wet or dry condition).
Hi, It is bettere to transfer after the running, for the problem Dominique explained. However, on website, you can find some protocol as put at 4°C and leave between glasses or store at 4°C without glasses in transfer buffer, or freeze in the fridge.
I think you have to try if they are suitable for you
Ideally you can keep the gel 15-30 in transfer buffer(Towbin). If you keep for longer there may be chances of disappearing of protein bands since Towbin buffer contain 20% methobol.
after you run the SDS-PAGE, you can leave the gel within the western-blot transfer buffer with methanol and acetic acid at 4ºc., before you run the western-blot
if u have not loaded the protein sample u may keep it for 1 day at 4 degree. but if u hv loaded the sample its better to run in order to avoid diffusion.
It is preferable that the transferring of protein bands exactly after the running of them to prevent diffusion of proteins specially in non purified proteins. Although, you can keep the gel up to 12 hours at 4◦ C.
I strictly recommend to make the transfer immediately after the SDS-PAGE in order to prevent diffusion of the proteins. Block the membrane with skim milk afterwards and then you can store the membrane at 4 degrees (in fridge)
Of course, store the membrane in TBS-T. Never leave the membrane dry!
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