Wat is the difference between native PAGE & SDS-PAGE?

Question

I want details and difference about that 2 electrophoresis technique and its uses.

Answer 1

In SDS-PAGE complexes are separated to their subunits, proteins are denatured and covered by SDS molecules at a ratio of approximately 1 SDS molecule per 2 amino acids. Thus any charge that the protein might have is masked by he huge negative charge by the SDS molecules and migration and thus separation of proteins depends mainly on their size.
That's why SDS page is commonly used for determing approximate molecular weight of proteins, for following the progress of protein purification, etc.


In native PAGE proteins retain their natural fold and can remain in complex. So the migration depends on the charge of the protein, the size, shape and if it is in complex with other molecules or if it oligomerizes.
For a example a protein that forms tetramers will give one band in an SDS-PAGE that corresponds to the monomer (provided that denaturation is complete) while on a native PAGE it can give more than one band, depending on the amount of each species (monomer, dimer, trimer, tetramer)
From native PAGE usually in combination with other techniques you can see the oligomerization state of your protein or study complexation reactions like protein-DNA (band-shift assays)

Answer 2

Native Page

Answer 3

Well, they say western is better for beginners because your kind of... closed in i guess. And you have a horn you can hold onto if needed. Also, you only use one hand to rein the horse. Also with Western horses, instead of a canter they 'lope' which is like a slower canter. While English, you are pushed up onto the horse more, so its not like your riding a saddle. With english, You also need a lot of leg muscle to hold on, but you will build that up over time. Some things you can do to start building it up are bike riding, swimming, wall sits, and other things like that :P Also with english you control the horse with both hands, and there is no horn that you can hold onto. Once you know english you can defiantly do western, because Western doesn't require quite as much leg, and only needs one hand! You can ride any style with any saddle, really. Sometimes I ride on a western saddle but the bit is english. Once your legs are strong enough you should be able to do bareback, too. You probably already know this, but just making sure so you understand if your horse spooks or something. Horses are prey animals and with the 'fight or flight' thing, they are made more to get away from the danger not fight it. So if they see something coming toward them, even just a paper bag, (depending on the horse) it could scare them, because all they see is something running at them. Good luck with your first horse! Btw im so jelous of where you live! lol

Answer 4

In Native PAGE,no denaturing agents are used to denature the protein sample.same buffers are used in all the three:
> the buffer,
> the gel and
>in the sample.
A single gel system is used to seperate the proteins based on their molecular weight.
Using this technique,enzymes can be seperated without denaturing them as they lose their activity upon denaturation.

In SDS-PAGE, an anionic detergent,SDS is used to, denature the protein sample and this sample is subjected to electrophoresis.

two different gel types are used:
stacking gel with pH-6.8 and a seperating gel of pH-8.8.
The buffer also varies with the gel and the tank.
The proteins get stacked between the glycine ions of tank buffer and chloride ions of buffer used to dissolve the gel in the stacking gel.
In seperating gel,the glycine and chloride ions move faster,and the proteins form bands according to their molecular weight.
This is the most widely used technique for the seperation of proteins and we can determine their molecular weight by comparing the bands with the marker protein sample run aside.