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 Cu(2+) in the BCA Kit's reagent B (4% cupric sulfate) in a manner similar to that of the protein.
authors.affiliation: NUSNNI-NanoCore, National University of Singapore, Singapore 117580, Singapore.
authors.forename: Sandeep Kumar
authors.initials: SK
authors.lastname: Vashist
authors.forename: BinBin
authors.initials: B
authors.lastname: Zhang
authors.forename: Dan
authors.initials: D
authors.lastname: Zheng
authors.forename: Khalid
authors.initials: K
authors.lastname: Al-Rubeaan
authors.forename: John H T
authors.initials: JH
authors.lastname: Luong
authors.forename: Fwu-Shan
authors.initials: FS
authors.lastname: Sheu
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002417:Cattle; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D011804:Quinolines / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000032:analysis*; D013388:Succinimides / Q000737:chemistry*
chemical_list: D011804:Quinolines; D013388:Succinimides; D012710:Serum Albumin, Bovine; C047117:bicinchoninic acid; C001426:N-hydroxysuccinimide; D005022:Ethyldimethylaminopropyl Carbodiimide