Drying Neupage gel - how to achieveing drying and prevent cracking (Jan/04/2009 )
I tried to dry a neupage gel (from Invitrogen), which was loaded with my protein samples. But the gel was not dried with the vacuum drier and turned to be cracked.
My protein samples were from tissue homogenates. A radio-labeled compound was incubated with the homogenates at 37 degree for 30 minutes. The reaction was stopped with ice-cold acetone. Then the precipitated proteins were washed with 1% SDS for three times. The precipitated proteins were dissolved in 1M NaOH and boiled with loading buffer and reducing reagent from the Invitrogen Neupage Gel Kit. After boiling, I neurtralized the samples with equal molar of HCl and loaded and ran the the samples on the Neupage gel.
I wanted to dry the gel after running for later exposure to the film. My goal is to detect any protein adducts formed by my compound. However, the drying step blocked me from moving on. I tried to fix the gel in 10% acetic acid and after that soaked the gel in 5% glycerol (a protocol I found online). But it didn't work.
The gel just cannot be dried.
Does anyone here can help with this problem?
Thank you very much!
We use the Tut's tomb apparatus to dry gels
(see this site: http://www.ideascientific.com/Tuts_tomb_inst.htm)
It's very simple, you could probably build the frame yourself. We would soak the gel and two pieces of cellophane in 20% ethanol then put the gel between the sheets of cellophane. The frame and bulldog clips would hold the whole thing taut at it dried to prevent cracking. It was also important to make sure there were no bauuble of air between the layer as this could also lead to cracking of the gel as it dries.