My 11% sds-pa gel broke into very many pieces during drying when the pump suddenly stopped working all of a sudden. is there any way to reconstitute the gel? It doesn't need to be pretty I just need to see my bands.
You actually did a pretty good job reassembling the gel, but why do you even bother drying them? In 4 years running SDS-PAGE I never dried a single gel. I simply scan them after staining/destaining and save the image. Much easier and less head-aching :)
No, you will not be able to even fix the 'band puzzle' correctly. hence no accurate inerpretation. Just let it go and run another, as much as it hurts.
If ur gel is not in very small piesces, u can add some water to swell the gel and then assamble it.u can see ur bands.
It is advisable to take pictures of all gels before attempting to dry on cellophane. You may try to reconstruct only to see the bands but may not be good to publish - and in most cases they are almost always useless after cracking. The best option is to run the gel again if you still had the samples.
I agree with Otieno.....try to solve the puzzle of gel pieces (not that difficult actually)....first stain- destain all the pieces...then assemble them and scan it!!
Many thanks. Actually swelling them with water and puzzling helped a lot.
Good job! I have reassembled a few gels, but I wouldn't have expected that yours could be saved.
Why dont u elute the product and run it again. Ifu hvnt sved the sample.....
You actually did a pretty good job reassembling the gel, but why do you even bother drying them? In 4 years running SDS-PAGE I never dried a single gel. I simply scan them after staining/destaining and save the image. Much easier and less head-aching :)
Well done- a good reconstruction. Now run the fresh samples and see if your re-creation was worth it. The reconstruction may have shifted some bands slightly, but what is important is that you have you darling bands. I hope this was not your masterpiece.
Sorry i forgot to say that my samples are radiolabeled. Blue bands are just antibodies (in this case they helped me puzzle the pieces back together). I have very small amount of protein therefore eluting would not help. I need to dry the gels in order to be able to expose them to a radiography screen. Wet gels don't really work. i also noticed that after drying, the pieces got a little streched and fit better than the wet state.
Very many thanks for your input, good to know you're not alone.
OK, you really need the gels dried for further use. I thought you only needed to save the gel to get a picture, hence my answer.
Good luck for when you expose the gel ;)
dear Zeynep I am sorry for your exp,I did many IP and it happens unfortunately. Next time I suggest to load onto the gel only half of the reactions, or try to look for a PhosphorImager in your Institute . Anyway, I hope you have used all precautions to avoid any contamination of you and your bench. As a tutor I should say that a gel in that state with radioactive samples must be thrown away, too dangerous to manipulate the pieces. Don't forget your health
Dear Paola thank you for reminding me about the health hazard. This was 35S. I took all the precautions necessary. But you are right, sometimes we get caught in the moment and forget about the safety.
Hi Zeynep,
to avoid breaking of your PA-Gel which should be used for autoradiography, you could try to cast your gels in the future with an inlay of NetFix from SERVA. We used this very frequently with radiolabeled 2D-Gels and could rescue 100% from breaking during the drying process.
Dear Murat,
Interesting, never heard of this. Will give it a try.
Tesekkürler :-)
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