Elsevier

Methods in Enzymology

Volume 233, 1994, Pages 240-250
Methods in Enzymology

[24] Nitric oxide assay using hemoglobin method

https://doi.org/10.1016/S0076-6879(94)33027-1Get rights and content

Publisher Summary

Hemoglobin, along with related hemoproteins such as myoglobin, can be used for the measurement of nitric oxide (NO). The technique is based on the direct reaction between NO and the oxygenated, ferrous form of hemoglobin (HbO2), which yields the ferric form, methemoglobin (metHb), and nitrate. This technique has the following advantages over other methods of NO measurement: all reagents are inexpensive and readily available, no complex techniques are required for sample collection or handling, the only large equipment needed for the basic technique is a spectrophotometer, and the assay is conducted under physiological conditions. This chapter discusses the basic method and describes the applications of the method to a variety of model systems, and points out possible interferences and artifacts. The chapter discusses the reaction that is the basis for the use of hemoproteins as efficient agents to block the biological effects of NO (the endothelium-derived relaxing factor, EDRF). This reaction is also proposed as a basis for the measurement of NO, using electron spin resonance (ESR) to detect and quantify the nitrosylhemoprotein product.

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