can gel be stored before transfer?

[wendyleye]

Hi guys,

Has anyone tried to store SDS-PAGEs for some time (e.g. 1 or 2 days) before electrotransferring? What are the conditions?

Any normally I use 40V o/n transfer, and it works great (in Biorad tank tranfer). So if I want to do a short time transfer instead, say 1 or 2 hr, should I increase the voltage? If so, rough how much?

Thank you.

[gandalfthegrey]

Hey
yes its possible to store SDS-PAGE gels easily there is:

Short term storage of SDS Gels

• completely seal the gel in a plastic bag (this prevents drying out)
• keep the gel at 4 degrees Celcius
• Storage like this is ok for 2-3 days
• The rate of protein diffusion is quite slow so usually gel definition of average sized proteins is maintained

Longer Storage of SDS Gels

• Start by completely sealing the gel in a plastic bag and freeze at minus 20 degrees Celcius
• SDS Gels can store for a month like this
• Western blots have no apparent loss of signal
  intensity or definition.

If you have very small proteins or you want to maintain extremely high resolution of two proteins which are at a similar molecular mass then use these storage methods with caution.

[admin]

Hey these are great tips thanks for posting.

You can also do this:

• Add cling wrap onto a small rigid support large enough to support your gel
• place your gel directly onto cling wrap
• Add a second sheet of cling wrap over the gel
• wrap the cling wrap around the gel and support snug
• Place into your -80 freezer.

NOTE:
your gel is fragile stored in this way.

Before freezing add gel into a small plastic box so nobody crushes it into 1 million pieces.

Thaw gel the next day wrapped, in luke warm water and then conduct western blot.

[anna.oskar]

I always do quick transfers.

Using the Bio-Rad setup: 120V
135 minutes
You must ice the tank

I've never had a problem doing it this way.