SDS-PAGE (polyacrylamide gel electrophoresis)

Animation showing the process of SDS-PAGE for the separation of proteins based on molecular weight.

Category:

Education

Tags:

• SDS-PAGE
• electrophoresis
• SDS
• polyacrylamide
• gel
• protein
• bromophenol
• blue
• dye
• pore
• buffer
• anode

[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.

[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.

[hamasaful]

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

[hamasaful]

@Rawrk92 that's on the agarose gel i think

[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!

[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.

[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'.

[MZee32]

Can anyone plz elaborate the function of the stacking layer ?