SDS-PAGE (polyacrylamide gel electrophoresis)

[mariaeatsbooks]

Great! :D

[Rawrk92]

I thought the negative charge came from the negatively charged oxygens in the phosphate groups of each DNA molecule?

What a headache!

[hamasaful]

@Rawrk92 that's on the agarose gel i think

[hamasaful]

@Rawrk92 agarose gel separates DNA but SDS PAGE and comassie blue , native page ( polyacrylamide gels ) are used to separate proteins

[MrTherandom]

how long do we keep the running buffer on?!?!?!

[shokokb1982]

Thank you very much for this video. I could understand SDS-PAGE. I'd like a video on the preparation of SDS-PAGE(ex dehydration of S-S bond, etc...)

[bjornb18]

1 PERSON IS LACKING A POWER SOURCE.

[MZee32]

Can anyone plz elaborate the function of the stacking layer ?

[generalgeert]

@MZee32

The stacking gel makes sure all proteins arrive at the resovling (or running) gel at the same time. This is important because if the proteins arrive at different times in the running layer they seperate not only on weight but also on who reaches the gel first! This is something you dont want. So in easy words: the function of the stacking gel is a 'fair' start. About how this fair start works google about 'Kohlrausch reactions'.

[aamiralius]

its too good explained

[nemokidd]

kool

[TarzanaGilbert]

Can this be run horizontally as well or just vertically as shown in animation? Thanks.

[Ferranax]

I've only ever done it horizontally. There's another gel electrophoresis video that shows horizontal migration.

[perebre]

maybe... it depends..

did you an electrophoresis for proteins??

I made a Northern Blot (RNA) and it was make in an horizontaly camara

[roborexasaurus]

@Ferranax

when I did it, I did it horozontally

[MrFedarov]

it possible

[Kistom22]

I dont know that you will read it but I do it vertically always. The agarose gel electrophoresis can you use horizontally

[dinkolino2]

why would any onewant to watch this? get a slave phd job and do it for real lol. Forget family and a normal life though

[hartistry1957]

What life?

[blindmice]

I laughed, I cried, wow what can I say - this video has it all. I want to lose my virginity to this video.

[hartistry1957]

I already have.

[eday84]

thank you for this video

[berlin45ers]

becuase small proteins experience less reistence when moving through the pores they move faser than the larger proteins----does that mean that the smaler proteins are nearer to the posetively charged anode?

[geneedinc]

It is true that the smaller proteins will experience less resistance and will travel further in the gel compared to larger proteins. And in SDS-PAGE, all proteins have a uniform negative charge because of the added SDS and migrate to the positively-charged anode.

[johndude969]

Just for clarification, is Ethidium Bromide used in this case to visualize the fragments or was something else used?

[geneedinc]

There are several ways to visualize the protein fragments in an SDS-Page gel, and we were not explicitly detailing which one was used in this animation. Coomassie blue is pretty much the most commonly used visualization stain, but silver staining is more sensitive. Fluorescent stains are also used. Because of the way Ethidium Bromide works (via intercalation of DNA base pairs), it really won't well for visualizing the proteins separated by an SDS-PAGE gel.

[johndude969]

Thank You

[Gigano1986]

I've actually used this method to determine the approximate mass of the enzyme vanillyl-alcohol oxidase in an experiment at my university. This video is very good representation of what happens.

[rax2099]

there is a variation for sds-page for enzymes.

[jabondeloza]

esta bueno el video... me sirve para estudiar :D