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protein extraction after cell lysing - (May/15/2009 )

i want to extract collagen from tissue implants. to measure its concentration later.

first i need to homogenize the tissue
then i need to lyse the cell and measure total protein content for nromalization of the collagen content i will measure later.

the questions are : 1- where is my protein (collagen) located after lysing ( in the pellet or in the supernatant ) ?
2- if collagen is in the supernatant how do i switch from the homogenzation solution (cell lysing buffer + protease inhibator ) to the exctraction solution (acetic acid + pepsin) . i was thinking by lyophelize ?!

thanks for help


Proteins (also known as polypeptides) are organic compounds made of amino acids arranged in a linear chain polymer and joined together by peptide bonds between the carboxyl and amino groups of adjacent amino acid residues.
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