Hello researchers
I just want to know what happen to the proteins molecules after they migrated out of the gel ( in SDS-PAGE) , of course the simple answer says that the protein migrated to the anode buffer then to the anode.
the question is what is the behavior of proteins molecule after they come in contact with the anode?
i will be greatfull for any help , and i will appreciate any article or review that discuss this behavior .
best regards .
Dear Hayder
The proteins will remain in the buffer more diluted and denaturated.
Best wishes
There is a lot of convection of the running buffer in the tank due to heating at the electrodes, so I don't think you can recover the protein from the anode. I think it will just be distributed in the running buffer throughout the tank. I've noticed that the bromphenol blue dye from the dye front floats upwards after being run off the gel and then dissipates.
If you want to try to recover the protein from SDS-PAGE, you can try electroelution.
http://www.bio-rad.com/en-us/category/preparative-electroelution-cells
Dear @Rosa Maria
the negatively charged molecules have to reach the anode because the buffer also electrically gradient . the question is , what is the behavior of this molecules after they reach the top of the electric gradient ( the anode ) ?
regards and thank you for this trying .
Dear Adam B . i have already noticed what you describe in your answer about the behavior of the dye , but theoretically the dye have to reach the anode due to its negative charge , so why it is ( the dye ) distributed in the buffer instead of keeping its migration toward the anode .
I think the reason is that there is a lot of heat generated at the electrodes. The buffer actually boils there. You can see the bubbles rising. The heat causes mixing of the running buffer due to convection, and that mixing overcomes the attraction between the anode and the dye.
Dear
Essentially after contact with the anode the proteins molecule are completely denatured
Marius
Thank Adam B Shapiro .
actually, the bubbles that we see near the electrodes are not due boiling , they formed because of gas formation ( mainly O2 and H2) due to Electrolysis of water . so i think there is an other reason to make the dye diffuse in the buffer.
please i need this answer , so if you have an other idea i will be grateful
regards ..
Dear Marius K. Somda
The protein molecules are already denatured by SDS and alkalinity, i don't care to be denatured or not , but i want to know the behavior of this molecule after they have entered the anode buffer .
Best regard .
Hi
The charged molecules "travel" toward the electrode, at some point, their density around the electrode reaches the limit where they kept away from the electrode because of the repulsion between them , this may be the cause why "the molecules do not reach the anode" in the way you think about!
dear Asim
well , maybe this is true .
But in this case we have to see a gradient concentration of charged molecule this gradient have to reach its maximal value near the anode , and we usually did not see this phenomena .
thank you . Michał Błażej Ponczek, for this detailed , logical and scientific answer.
I think this is the best answer
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