Measurement of heme concentration.
Heme (iron protoporphyrin IX) is a prosthetic group for a number of hemoproteins in different tissues (e.g., hemoglobin, myoglobin, cytochrome P-450s, mitochondrial cytochromes, catalases, and peroxidases). Mutations in the biosynthetic pathway can affect the synthesis and/or degradation of heme. Several assays are provided in this unit for quantifying heme: a spectrophotometric assay based on the characteristic absorption spectrum of oxidized and reduced form of the hemochrome formed by replacing the nitrogen ligands with pyridine; a fluorescence assay based on removal of the iron by a heated, strong oxalic acid solution to produce fluorescent protoporphyrin; a reversed-phase HPLC assay to measure heme and intermediates in the synthetic pathway; and a radiometric assay to measure newly synthesized heme in tissue culture cells.
Sinclair
P R
PR
Veterans Administration Medical Center, White River Junction, Vermont, USA.
Gorman
N
N
Jacobs
J M
JM
D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005453:Fluorescence; D006418:Heme / Q000032:analysis*; D006801:Humans; D010084:Oxidation-Reduction; D011874:Radiometry / Q000379:methods; D013053:Spectrophotometry / Q000379:methods
D006418:Heme