native page vs. non-reducing

[theonlytycrane]

I'm trying to make sense of any additional information that native page would provide over non-reducing? Would just running one of these + a reducing gel provide enough information for protein analysis?

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[salemstein]

If I'm understanding it correctly, native page doesn't alter anything on the protein. A in, A out. Non-reducing doesnt break any covalent bonds, and assuming its SDS + non-reducing, only H bonds are broken.

 

[betterfuture]

SDS + reducing means SDS with beta mercaptoethanol and SDS + nonreducing means SDS without beta mercaptoethanol, right?

 

[theonlytycrane]

@betterfuture yes.

 

[Astra]

Native page will give you information about protein structure.

Say you have 2 proteins of 100 kD.

You run them.

However you notice that one protein migrated a bit further than the other. This indicates that the protein that migrated further is more tightly folded which allowed a further migration.

This is the same logic that causes supercoiled DNA to migrate further than regular DNA.

 

[theonlytycrane]

@Astra118 ah this helps! But for a single protein, a native page wouldn't give any additional information than from a non-reducing page, correct?

 

[aldol16]

@Astra118 ah this helps! But for a single protein, a native page wouldn't give any additional information than from a non-reducing page, correct?

I think you mean monomeric protein. And a native PAGE would still give you a rough idea of whether the protein is globular or not because proteins with certain secondary/tertiary structures migrate in certain ways.