Biotechniques | Principles of SDS-PAGE (Protein Separation)

As always, the steps of an SDS-PAGE will depend on your specific lab and protocol. In this video, we will explore the general theory of separating and visualizing protein by SDS-PAGE, a technique very similar to DNA agarose gel electrophoresis. This procedure is the precursor to the Western blot.
Standard curve tutorial in next video:

[@bhumikamago]

Quick question Migration of the protein is dependent on 2 factors Its molecular weight And its charge So more the molecular weight less the distance it will cover But higher molecular weight also means more sds and more negatively charged so doesn’t it mean it should get more attracted towards anode ? So if the molecular weight is high it will travel more or less than other protein with less molecular weight and also less negative charge

[@moviesharer5542]

i tumbs up for you can directly speak out the professional words!

[@esabelroncon4761]

First off: amazing explanation - thank you so much! Now for my question: for the SDS-PAGE after you run electrophoresis, at 11:34 why are the four different lanes showing less and less proteins as you move across? are you performing some sort of washing step? If that is the case, how is it done? And furthermore, if by lane 4 you can see your protein of interest, why do a western blot?

[@kaushikilohiya2137]

This is a helpfull vedio

[@PreetiSharma-jt9tp]

Very helpful

[@l3ft.br4in]

super helpful, but does the sodium from sds do anything? like where does it go?

[@esabelroncon4761]

no it isnt really useful. it doesnt 'go' anywhere. it still exists like a spectator ion

[@samuelh545]

PERFECT