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What is different between Native protein and Denatured protein samples. - (May/08/2013 )

I started to learn western blot from Denatured protein technique.

What is the purpose to run sample with native protein ?

-Tai-

Do you mean you are also running native protein on the same gel as the denatured, or that you are running other native protein gels?

-bob1-

native protein will have the correct conformation, subunit structure, charge, size, etc.

denatured protein will give you subunit size.

some antibodies will only recognize denatured protein, some will only recognize native protein, and some will recognize both.

-mdfenko-

Thank you for your response!

bob1 on Wed May 8 21:57:45 2013 said:


Do you mean you are also running native protein on the same gel as the denatured, or that you are running other native proteins?

My question is about when should I design to run WB with the native protein?
and what is the different of the result of the native protein or the denatured protein from WB ?
or is there any meaning to interpret our results, if we use the native protein or the denatured protein?


mdfenko on Thu May 9 12:54:54 2013 said:


native protein will have the correct conformation, subunit structure, charge, size, etc.

denatured protein will give you subunit size.

some antibodies will only recognize denatured protein, some will only recognize native protein, and some will recognize both.


That mean, it depends on antibody that is available for our particular protein, right ?
Then, we can design to prepare the lysis buffer for native or denature sample.

-Tai-

Tai on Wed May 15 08:24:42 2013 said:


That mean, it depends on antibody that is available for our particular protein, right ?
Then, we can design to prepare the lysis buffer for native or denature sample.

it also depends on what feature of the protein you want to show

-mdfenko-