In my current western blot routine, I cast my (10%) gels on day 1 and store them at 4 degrees Celsius in soaked paper and cling wrap overnight.
I intend to use materials as freshly as possible if time allows it, but I want to gain flexibility if this is necessary during my project.
Thank you in advance,
IJsbrand
I usually fill the wells with buffer/leave a comb in, then wrap them in a soaking wet paper towel, saran wrap, and then foil and keep them at 4C. I've had them work weeks, probably a month or more. If the stacking gel is starting to shrivel or dry out, or if the paper towels are drying out thats a sign for me that its been too long.
Thank you for your answers. I guess a week should be fine then? I'll do some experiments to investigate. Would the protein transfers also be affected by the age of the gel or only the electrophoresis?
The longer you store in the fridge, the more the gel will lose its humidity, i.e. dried up. Therefore, the percentage of your SDS gel will be affected if its water content has evaporated. Based on this understanding, the electrophoresis will be affected as smaller protein sizes are now not able to run down the gel.
The best way is to put it in a zip lock bag with water and you can store it at 4C up to a month, a word of caution is that as time passes the lower MW bands will be a bit fuzzier if that makes sense when you run your gel.
I usually fill the wells with buffer/leave a comb in, then wrap them in a soaking wet paper towel, saran wrap, and then foil and keep them at 4C. I've had them work weeks, probably a month or more. If the stacking gel is starting to shrivel or dry out, or if the paper towels are drying out thats a sign for me that its been too long.
I have used gels that are 4-6 weeks old with no problems :) As long as it has been kept moist, at 4 degrees (I usually wrap in wet paper towel, then in tinfoil/ cling wrap). Just remember to check the wells when unwrapping the gel- i they show any signs of drying out/ deformities then do not use.
Agree with most answers suggested, as long the moisture is maintained by keeping the gel in air tight bags it can survive for 6 > 8 weeks. Try to follow the storing conditions given by ready made gel producers. Addition of 2 to5% glycerol in the storing medium can help in maintaining the moisture.
Its depends upon how you stored your gels. If we used wet tissues at 4°C then it will work well almost 15 days.
We used to prepare just running gel and that could be stored in wet tissue paper in running buffer at 4degC for weeks. Stacking gel was prepared fresh. I also used to store the whole gel, but I left the comb inside. However, that one I usually used up to 2 weeks, so I never checked after.
If gels are stored for too long, they may not only dry out, but diffusion may equilibrate the buffers in stacking and resolving gel, so that proteins don't stack properly anymore, leading to fuzzy bands. In a gel, and in the cold, diffusion is slowed, so this process takes time. I make a point to use my gels within 2 weeks, and never had a problem.
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