Simplified extraction methods for the rapid determination of species content of adipose tissue based on the detection of tni in immunoassays

Abstract

A method for extracting protein from a fat-containing sample is described comprising the steps of separating solidified fat and solid residues from a centrifuged mixture of protein, fat, other solid materials and aqueous solution of phosphate buffered saline to form an aqueous phase containing the protein. The aqueous phase is then filtered through a filter to separate a clear protein extract from the mixture.

1 . (canceled) 
     
     
         2 . A method comprising the following steps:
 (a) separating solidified fat and other solid materials from a mixture to thereby form an aqueous phase containing soluble proteins, and   (b) filtering the aqueous phase through a filter to thereby separate a protein extract from the mixture,   wherein the mixture comprises an aqueous phase containing a phosphate buffered saline and containing the soluble proteins and insoluble and/or immiscible substances from a pre-warmed and ground fat-containing sample,   wherein the method comprises conducting the following step before step (a):   (c) centrifuging the mixture at a reduced temperature.   
     
     
         3 . The method of  claim 2 , wherein the reduced temperature is in a range from 1 to 10° C. 
     
     
         4 . The method of  claim 2 , wherein step (c) is conducted for 15 to 60 minutes at speed sufficient to form a fat layer comprising the solidified fat, a solid tissue residue, and a liquid layer comprising the aqueous phase containing the phosphate buffered saline and the soluble proteins, and wherein step (a) comprises separating the fat layer from the liquid layer. 
     
     
         5 . The method of  claim 2 , wherein the method comprises conducting the following step before step (c):
 (d) forming the mixture by mixing a softened ground fat-containing sample with the phosphate buffered saline.   
     
     
         6 . The method of  claim 5 , wherein step (c) is performed after the mixture has been allowed to sit for at least 1 hour at room temperature after step (d). 
     
     
         7 . The method of  claim 5 , wherein the method comprises the following step:
 (e) forming the softened ground fat-containing sample by heating a ground fat-containing sample in an oven for 30 to 60 minutes at an oven heating temperature of between 60° C. and 80° C. to thereby soften the ground fat-containing sample.   
     
     
         8 . (canceled) 
     
     
         9 . (canceled)