Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay |
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| Authors: | Sandeep Kumar Vashist BinBin Zhang Dan Zheng Khalid Al-Rubeaan John H.T. Luong Fwu-Shan Sheu |
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| Affiliation: | aNUSNNI-NanoCore, National University of Singapore, T-Lab Level 11, 5A Engineering Drive 1, Singapore 117580, Singapore;bDepartment of Electrical and Computer Engineering, National University of Singapore, 5A Engineering Drive 1, Singapore 117576, Singapore;cDepartment of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore;dDepartment of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore;eUniversity Diabetes Center, King Saud University, P.O. Box 18397, Riyadh 11415, Saudi Arabia;fBiotechnology Research Institute, National Research Council Canada, Montreal, Quebec, Canada H4P 2R2 |
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| 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. |
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