Pyridine Hemochromagen Assay for Determining the Concentration of Heme in Purified Protein Solutions
Department
Natural Sciences and Mathematics
Document Type
Article
Source
Bio-Protocol
Publication Date
9-20-2015
Volume
5
Issue
18
Abstract
Heme is a common cofactor in proteins, found in hemoglobin, myoglobin, cytochrome P450, DGCR8, and nitric oxide synthase, among others. This protocol describes a method for quantifying heme that works best in purified protein samples. This protocol might be used to, for example, determine whether a given heme-binding protein is fully occupied by heme, thus allowing correlation of heme content with activity. This requires the absolute heme concentration and an accurate protein concentration. Another use is to determine the extinction coefficients of a heme-bound protein. This assay is fast, easy, and reproducible if done correctly.
PubMed ID
27390766
Rights
Copyright: © 2015 The Authors; exclusive licensee Bio-protocol LLC.