Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr 30 min
- Sensitivity: 0.015 µM
Product nameProtein Carbonyl Content Assay Kit
See all Protein Carbonyl kits
Assay time1h 30m
Protein Carbonyl Content Assay Kit ab126287 is designed to provide a simple and accurate method of quantifying carbonyls in protein samples.
The protein carbonl assay protocol is based on the reaction of DNPH with protein carbonyls. This is the most popular assay type for measuring protein carbonyl content. DNP hydrazones formed in this reaction are easily quantifiable at 375 nm absorbance.
When used with, for example a 1 mg (~15 nmol) sample, this protein carbonyl assay protocol has a detection limit of about 0.15 nmol carbonyl. For context, BSA typically contains approximately 1-3 nmol carbonyl/mg.
Protein carbonyl assay protocol summary:
- add DNPH to samples and incubate for 10 min
- add TCA to samples, incubate for 5 min, spin for 2 min, and discard supernatant
- wash pellet by sonicating in acetone, spin and remove acetone, repeat acetone wash step
- add Guanidine solution and resolubilize pellet, and transfer to plate
- analyze with a microplate reader to measure protein carbonyl content, also perform a protein assay (eg. BCA assay) on samples to establish protein concentration
This is the most popular Protein Carbonyl Content assay kit. Other assays include:
- Protein Carbonyl Content Assay (Fluorometric) ab235631
- Protein Carbonyl Content ELISA (DNPH) ab238536
- Protein Carbonyl Content Western Blot Assay ab178020
Review the oxidative stress marker and assay guide, or the full metabolism assay guide to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.
Storage instructionsStore at +4°C. Please refer to protocols.
Components Identifier 100 tests 10% Streptozocin Solution Blue 1 x 1ml 87% TCA Solution NM 1 x 3ml 6 M Guanidine Solution WM 1 x 20ml 96-Well Clear Plate 1 unit DNPH Solution Amber 1 x 11ml
Typical Standard Curve using BCA Protein Quantification Assay (ab102536)
Representative Data Obtained Using the Protein Carbonyl Content Assay Kit
ab126287 has been referenced in 25 publications.
- Lei K et al. C/EBPß mediates NQO1 and GSTP1 anti-oxidative reductases expression in glioblastoma, promoting brain tumor proliferation. Redox Biol 34:101578 (2020). PubMed: 32526700
- Lei K et al. Baicalin Represses C/EBPß via Its Antioxidative Effect in Parkinson's Disease. Oxid Med Cell Longev 2020:8951907 (2020). PubMed: 32566108
- Yen CJ et al. Effect of Exercise Training on Exercise Tolerance and Level of Oxidative Stress for Head and Neck Cancer Patients Following Chemotherapy. Front Oncol 10:1536 (2020). PubMed: 33014797
- Lei K et al. Discovery of a dual inhibitor of NQO1 and GSTP1 for treating glioblastoma. J Hematol Oncol 13:141 (2020). PubMed: 33087132
- Pusic KM et al. IFN?-Stimulated Dendritic Cell Exosomes for Treatment of Migraine Modeled Using Spreading Depression. Front Neurosci 13:942 (2019). PubMed: 31551696