Title

Pyridine Hemochromagen Assay for Determining the Concentration of Heme in Purified Protein Solutions

Digital Object Identifier / DOI

https://doi.org/10.21769/bioprotoc.1594

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.

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