Determination of protein carbonyls in plasma, cell extracts, tissue homogenates, isolated proteins: Focus on sample preparation and derivatization conditions

TY - JOUR

T1 - Determination of protein carbonyls in plasma, cell extracts, tissue homogenates, isolated proteins

T2 - Focus on sample preparation and derivatization conditions

AU - Weber, Daniela

AU - Davies, Michael J.

AU - Grune, Tilman

PY - 2015/8/1

Y1 - 2015/8/1

N2 - Protein oxidation is involved in regulatory physiological events as well as in damage to tissues and is thought to play a key role in the pathophysiology of diseases and in the aging process. Protein-bound carbonyls represent a marker of global protein oxidation, as they are generated by multiple different reactive oxygen species in blood, tissues and cells. Sample preparation and stabilization are key steps in the accurate quantification of oxidation-related products and examination of physiological/pathological processes. This review therefore focuses on the sample preparation processes used in the most relevant methods to detect protein carbonyls after derivatization with 2,4-dinitrophenylhydrazine with an emphasis on measurement in plasma, cells, organ homogenates, isolated proteins and organelles. Sample preparation, derivatization conditions and protein handling are presented for the spectrophotometric and HPLC method as well as for immunoblotting and ELISA. An extensive overview covering these methods in previously published articles is given for researchers who plan to measure protein carbonyls in different samples.

AB - Protein oxidation is involved in regulatory physiological events as well as in damage to tissues and is thought to play a key role in the pathophysiology of diseases and in the aging process. Protein-bound carbonyls represent a marker of global protein oxidation, as they are generated by multiple different reactive oxygen species in blood, tissues and cells. Sample preparation and stabilization are key steps in the accurate quantification of oxidation-related products and examination of physiological/pathological processes. This review therefore focuses on the sample preparation processes used in the most relevant methods to detect protein carbonyls after derivatization with 2,4-dinitrophenylhydrazine with an emphasis on measurement in plasma, cells, organ homogenates, isolated proteins and organelles. Sample preparation, derivatization conditions and protein handling are presented for the spectrophotometric and HPLC method as well as for immunoblotting and ELISA. An extensive overview covering these methods in previously published articles is given for researchers who plan to measure protein carbonyls in different samples.

KW - 2,4-Dinitrophenylhydrazine

KW - ELISA

KW - Immunoblotting

KW - Protein carbonyls

KW - Sample preparation

KW - Spectrophotometry

U2 - 10.1016/j.redox.2015.06.005

DO - 10.1016/j.redox.2015.06.005

M3 - Review

C2 - 26141921

AN - SCOPUS:84933520752

SN - 2213-2317

VL - 5

SP - 367

EP - 380

JO - Redox Biology

JF - Redox Biology

ER -