SDS PAGE | Stacking vs Resolving gel

SDS PAGE is a common technique used for the analysis of proteins. The SDS PAGE gel is a discontinuous gel. The stacking gel has a pH of 6.8 where as the resolving gel has pH of 8.8. The stacking gel plays an important role in stacking all protein molecules in one line so that excellent resolution can be obtained in the resolving gel.

[@vivianwkli]

I think the Cl- moves faster and should be the leading ion, then the protein, with negatively charged glycine at the top of the sandwich. Otherwise it explains the SDS-page stacking and resolving gel nicely. Thank you.

[@bobu5213]

But this wouldn't make any sense, if the glycine has a zero net charge but is at the top of the sandwich, there is no force pushing the proteins down into a sandwich. The glycine isn't pulled anywhere due to the zero net charge. In the video's example, the proteins push the glycine which creates resistance as the glycine doesn't move on it's own as the non-charged variant.

[@rolandgromes636]

@@bobu5213 Glycine has an IP of 5.97, so at pH 6.8 it is negatively charged, not neutral. It's just that the glycine is more frequently in the neutral state and therefore moving slower. Glycine forming a "wall" makes no sense at all UNLESS it is negatively charged and therefore repells the equally negatively charged proteins. What actually happens is that the fast chloride and the slow glycine leave the proteins inbetween in a region of low ion concentration, resulting in them being the only carriers of current, i.e. getting accelerated until they run into the strongly charged chloride front

[@taylork2874]

Great video!

[@nidhis1289]

Thank you so much!

[@indiabiologyexplains5013]

I have a question. After the sandwich of chlorine ions, proteins, and glycine reaches the resolving gel, the glycine leaves for the positive electrode. Do the Chlorine ions remain in the gel with the traveling proteins, or are they removed by some other process.

[@estebangodoy2576]

I couldn't understand this, you saved me, thank you very much.

[@theobroma5179]

What's the best percentage of separation and stacking gel for protein around 37 kda?

[@sebmilani8657]

Perfectly explained

[@DashRJ2000]

Thank you sir 😊

[@andrewrodriguez4516]

Thank you sir

[@rinitta5214]

Sir can you please upload starch gel electrophoresis

[@Sid-ro2ye]

Hi there,
great video.
Do the proteins automatically enter the resolving gel (even without changing the voltage) or do they only enter into the resolving gel the moment the voltage is changed?
Thanks!

[@janisisters6613]

Wonderful explanation 😊😊

[@dilaraozturk1896]

You stated that glycine and Cl ions are in the buffer. Could this buffer be a Tris buffer? Which product in polyacrylamide gel contains glycine and chlorine?

[@noflexzone2.055]

Your accent sounds much more Western compared to your previous videos... two thumbs up :)

[@sahla2943]

Thank you

[@martynaszarakauskas3105]

actually, amazing video

[@ezra47986]

woooww thank you

[@raniveniindu2804]

Sir please upload bioreactors

[@timurmakarov9769]

БАЗА

[@amrutagore1884]

Asmw