protein quantification

[aftabac]

Hi forum fellows
what is the best way to quantify the unknown protien sample, the sample may be a single purified protien or complex mixtures of proteins.


aftab

[kiki06]

Difficult question. If you want to know the general amount of protein in your sample you could use a BIORAD protein quantification assay. You could also do a UV spec analysis of your protein sample.

not sure how you could determine your exact protein amount.... do you have an antibody for that protein??? if so maybe you could do an ELISA assay...

just trying to help

[MicroKiller]

You can try either Bradford or Lowry method.. u can check it up online.. this is for quantitation.. but if u wnt to do determination... if u know wht the protein is.. elisa, blotting may be a good idea...

[kalaig]

Well! If you really want it perfect, digest hydrolyze proteins (alkali+autoclaving) into individual amino acids and estimate the amino acids.

[moleculardude]

Hey Tuttu and Kalaig!!

welcome to the forums.... really nice posts thanks for the information

[tuttu]

Hi
I think its a late answer anyway, u may probably first run a SDS-PAGE and check the protein profile of ur sample, and if its a purified one, u will mostly get a single band(may vary based on subunit trouble) and by a bradford u can estimate it. Well if its a mixture u may still quantify by bradford the total protein conc.
Hope its of use!!

[tuttu]

Hi moleculardude thanks for the welcome...

[songsong]

I agree with tuttu. It is the best idea for purfied protein.