Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay

Sandeep Kumar Vashist; Binbin Zhang; Dan Zheng; Khalid Al-Rubeaan; John H.T. Luong; Fwu Shan Sheu

doi:10.1016/j.ab.2011.05.045

Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay

Sandeep Kumar Vashist
, Binbin Zhang
, Dan Zheng
, Khalid Al-Rubeaan
, John H.T. Luong
, Fwu Shan Sheu

National University of Singapore
King Saud University
National Research Council of Canada

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

Abstract

This study revealed a major interference from sulfo-N-hydroxysuccinimide (sulfo-NHS) in the bicinchoninic acid (BCA) protein assay. Sulfo-NHS, a common reagent used in bioconjugation and analytical biochemistry, exhibited absorbance signals and absorbance peaks at 562 nm, comparable to bovine serum albumin (BSA). However, the combined absorbance of sulfo-NHS and BSA was not strictly additive. The sulfo-NHS interference was suggested to be caused by the reduction of Cu²⁺ in the BCA Kit's reagent B (4% cupric sulfate) in a manner similar to that of the protein. Crown

Original language	English
Pages (from-to)	156-158
Number of pages	3
Journal	Analytical Biochemistry
Volume	417
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2011.05.045
Publication status	Published - Oct 2011
Externally published	Yes

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10.1016/j.ab.2011.05.045

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title = "Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay",

abstract = "This study revealed a major interference from sulfo-N-hydroxysuccinimide (sulfo-NHS) in the bicinchoninic acid (BCA) protein assay. Sulfo-NHS, a common reagent used in bioconjugation and analytical biochemistry, exhibited absorbance signals and absorbance peaks at 562 nm, comparable to bovine serum albumin (BSA). However, the combined absorbance of sulfo-NHS and BSA was not strictly additive. The sulfo-NHS interference was suggested to be caused by the reduction of Cu2+ in the BCA Kit's reagent B (4\% cupric sulfate) in a manner similar to that of the protein. Crown",

author = "Vashist, \{Sandeep Kumar\} and Binbin Zhang and Dan Zheng and Khalid Al-Rubeaan and Luong, \{John H.T.\} and Sheu, \{Fwu Shan\}",

year = "2011",

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doi = "10.1016/j.ab.2011.05.045",

language = "English",

volume = "417",

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AU - Vashist, Sandeep Kumar

AU - Zhang, Binbin

AU - Zheng, Dan

AU - Al-Rubeaan, Khalid

AU - Luong, John H.T.

AU - Sheu, Fwu Shan

PY - 2011/10

Y1 - 2011/10

N2 - This study revealed a major interference from sulfo-N-hydroxysuccinimide (sulfo-NHS) in the bicinchoninic acid (BCA) protein assay. Sulfo-NHS, a common reagent used in bioconjugation and analytical biochemistry, exhibited absorbance signals and absorbance peaks at 562 nm, comparable to bovine serum albumin (BSA). However, the combined absorbance of sulfo-NHS and BSA was not strictly additive. The sulfo-NHS interference was suggested to be caused by the reduction of Cu2+ in the BCA Kit's reagent B (4% cupric sulfate) in a manner similar to that of the protein. Crown

AB - This study revealed a major interference from sulfo-N-hydroxysuccinimide (sulfo-NHS) in the bicinchoninic acid (BCA) protein assay. Sulfo-NHS, a common reagent used in bioconjugation and analytical biochemistry, exhibited absorbance signals and absorbance peaks at 562 nm, comparable to bovine serum albumin (BSA). However, the combined absorbance of sulfo-NHS and BSA was not strictly additive. The sulfo-NHS interference was suggested to be caused by the reduction of Cu2+ in the BCA Kit's reagent B (4% cupric sulfate) in a manner similar to that of the protein. Crown

UR - https://www.scopus.com/pages/publications/80655148685

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M3 - Article

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AN - SCOPUS:80655148685

SN - 0003-2697

VL - 417

SP - 156

EP - 158

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 1

ER -

Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay

Abstract

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