Binding of EDTA to DEAE-cellulose and its interference with protein determinations

W. W. Ward; R. J. Fastiggi

doi:10.1016/0003-2697(72)90494-0

Binding of EDTA to DEAE-cellulose and its interference with protein determinations

W. W. Ward, R. J. Fastiggi

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

EDTA binds to DEAE-cellulose columns and salt-eluted fractions may contain as much as 20 times the original concentration of EDTA. If not corrected for, this high concentration of EDTA will interfere with three of the most frequently used protein assays. The errors encountered were reductions in apparent protein concentration of 50% as measured by the Lowry assay, 25% by the biuret determination, and as much as 90% by the spectrophotometric method. The mechanisms for these interferences appear to be chelation of copper by EDTA, in the case of the Lowry and biuret assays, and differential absorption of ultraviolet light by EDTA in the spectrophotometric case. Several possible methods for overcoming these interferences are discussed.

Original language	English (US)
Pages (from-to)	154-162
Number of pages	9
Journal	Analytical Biochemistry
Volume	50
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(72)90494-0
State	Published - Nov 1972
Externally published	Yes

All Science Journal Classification (ASJC) codes

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0003-2697(72)90494-0

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title = "Binding of EDTA to DEAE-cellulose and its interference with protein determinations",

abstract = "EDTA binds to DEAE-cellulose columns and salt-eluted fractions may contain as much as 20 times the original concentration of EDTA. If not corrected for, this high concentration of EDTA will interfere with three of the most frequently used protein assays. The errors encountered were reductions in apparent protein concentration of 50% as measured by the Lowry assay, 25% by the biuret determination, and as much as 90% by the spectrophotometric method. The mechanisms for these interferences appear to be chelation of copper by EDTA, in the case of the Lowry and biuret assays, and differential absorption of ultraviolet light by EDTA in the spectrophotometric case. Several possible methods for overcoming these interferences are discussed.",

author = "Ward, {W. W.} and Fastiggi, {R. J.}",

note = "Funding Information: This work was supported by Atomic Energy Commission grant AEC (11-l) 3277 (to H. H. S.). One of us (W. W. W.) is the recipient of a National Science Foundation Graduate Fellowship.",

year = "1972",

month = nov,

doi = "10.1016/0003-2697(72)90494-0",

language = "English (US)",

volume = "50",

pages = "154--162",

journal = "Analytical Biochemistry",

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T1 - Binding of EDTA to DEAE-cellulose and its interference with protein determinations

AU - Ward, W. W.

AU - Fastiggi, R. J.

N1 - Funding Information: This work was supported by Atomic Energy Commission grant AEC (11-l) 3277 (to H. H. S.). One of us (W. W. W.) is the recipient of a National Science Foundation Graduate Fellowship.

PY - 1972/11

Y1 - 1972/11

N2 - EDTA binds to DEAE-cellulose columns and salt-eluted fractions may contain as much as 20 times the original concentration of EDTA. If not corrected for, this high concentration of EDTA will interfere with three of the most frequently used protein assays. The errors encountered were reductions in apparent protein concentration of 50% as measured by the Lowry assay, 25% by the biuret determination, and as much as 90% by the spectrophotometric method. The mechanisms for these interferences appear to be chelation of copper by EDTA, in the case of the Lowry and biuret assays, and differential absorption of ultraviolet light by EDTA in the spectrophotometric case. Several possible methods for overcoming these interferences are discussed.

AB - EDTA binds to DEAE-cellulose columns and salt-eluted fractions may contain as much as 20 times the original concentration of EDTA. If not corrected for, this high concentration of EDTA will interfere with three of the most frequently used protein assays. The errors encountered were reductions in apparent protein concentration of 50% as measured by the Lowry assay, 25% by the biuret determination, and as much as 90% by the spectrophotometric method. The mechanisms for these interferences appear to be chelation of copper by EDTA, in the case of the Lowry and biuret assays, and differential absorption of ultraviolet light by EDTA in the spectrophotometric case. Several possible methods for overcoming these interferences are discussed.

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JO - Analytical Biochemistry

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ER -

Binding of EDTA to DEAE-cellulose and its interference with protein determinations

Abstract

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