I tried to separate casein from whey proteins by isoelectric precipitation (pH 4.6) using 10% HCl. Then, with the whey fraction, I tried to separate lactalbumin from lactoglobulin with different percentages of ammonium sulfate precipitation (30%,50% and 100%).
As you can see in the SDS-PAGE gel, casein is present in whey fraction and "contaminated" the other fractions.
Also, on ladder 3 we can see lactalbumin and lactoglobulin that was not expected with isoelectric precipitation. However, it was recently reported that β-lactoglobulin and α-lactalbumin were identified in bovine casein fractions by 2-DE combined with MS approach. This could justify the presence of lactalbumin and lactoglobulin on the casein fraction.
Can anyone tell me how to get rid of casein from whey fraction?
I'm using skimmed milk from cow.
I did that. I precipitate casein with 10%HCl until pH 4.6, then centrifuge 9000rpm, 15min. The pellet is casein and the supernantant the whey fraction.
But the whey fraction is contaminated with casein (SDS-PAGE - ladder 2)
I read that after collecting the whey fraction, it should be centrifuge to remove casein remains.
I will try either centrifuge or filter the whey fraction.
Thanks
Go through this research publication by Joseph Dumpler et al. You will see a detailed procedure for casein isolation/precipitation
Separate the casein from the serum
The separation is carried out using acid, alcohol, enzyme, bacteria, salt
You can use the acid to separate casein from milk serum Proteins.
Prof. Dr. Shehadeh Kaskous
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