TY - JOUR
T1 - Sensitive protein assay in presence of high levels of lipid
AU - Kaplan, Ronald S.
AU - Pedersen, Peter L.
N1 - Funding Information:
This work was supported by National Science FoundationG rant PCM 8300772 to PLP.
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 1989/1/1
Y1 - 1989/1/1
N2 - This chapter describes a method that is capable of accurately measuring low amounts of a protein in the presence of very high levels of lipid. This procedure was developed from the amido black 10 B methods of Schaffner and Weissmann and Newman et al. and incorporates several critical modifications that enable an assay to be performed with lipid-containing samples without any interference. One approach has been to remove an interfering lipid by extraction with organic solvents. However, because certain proteins display a limited solubility in such solvents, this strategy often fails. Another widely used approach involves the inclusion of sodium dodecyl sulfate (SDS) in a modified Lowry procedure to reduce lipid (and detergent) interference. As oxidized lipid continues to react to produce a substantial amount of color in the Lowry assay and as most lipid samples are partially oxidized, this procedure is not suitable for the accurate measurements of a protein in samples containing excess of lipid.
AB - This chapter describes a method that is capable of accurately measuring low amounts of a protein in the presence of very high levels of lipid. This procedure was developed from the amido black 10 B methods of Schaffner and Weissmann and Newman et al. and incorporates several critical modifications that enable an assay to be performed with lipid-containing samples without any interference. One approach has been to remove an interfering lipid by extraction with organic solvents. However, because certain proteins display a limited solubility in such solvents, this strategy often fails. Another widely used approach involves the inclusion of sodium dodecyl sulfate (SDS) in a modified Lowry procedure to reduce lipid (and detergent) interference. As oxidized lipid continues to react to produce a substantial amount of color in the Lowry assay and as most lipid samples are partially oxidized, this procedure is not suitable for the accurate measurements of a protein in samples containing excess of lipid.
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U2 - 10.1016/S0076-6879(89)72025-5
DO - 10.1016/S0076-6879(89)72025-5
M3 - Article
C2 - 2747536
AN - SCOPUS:0024362028
SN - 0076-6879
VL - 172
SP - 393
EP - 399
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -