蛋白Carbonyl Assay试剂盒(Western Blot)

产品详情

Protein Carbonyl Assay Kit (Western Blot) ab178020 is designed for the measurement of protein carbonyl groups that are created by the oxidation of proteins. This can be through reaction with ozone or oxides of nitrogen, or by metal catalyzed oxidation.

In the protein carbonyl assay protocol, carbonyl groups in protein side chains are derivatized to DNP-hydrazone by reaction with DNPH. The DNP moieties are detected using an anti-DNP antibody and western blotting.

Using an immunoblotting method for the protein carbonyl assay has the advantage that individual oxidized proteins are separated and identified from a complex mixture by SDS-PAGE. The oxidative status of each protein can be compared between samples.

Protein carbonyl assay protocol summary:
- extract proteins from samples with extraction buffer, 20 min incubation, centrifugation at 18,000 x g for 20 min, and retention of the supernatant
- denature proteins with SDS solution
- incubate with DNPH for 15 min
- add neutralization solution
- run samples on polyacrylamide gel
- transfer proteins to membrane
- block for 1 hr
- incubate with DNP antibody for 3 hr, and wash
- incubate with HRP goat anti-rabbit secondary antibody for 1 hr, and wash
- develop with ECL and acquire image

Note: RIPA buffer is not compatible with ab178020

Previously called Oxidized Protein Assay Kit (Western blot). Oxygen-derived free radicals have been implicated in important roles in aging, apoptosis, and cancer. These highly reactive chemical species are also involved in a wide variety of clinical disorders, such as atherosclerosis, cataractogenesis, neurodegenerative diseases, chronic inflammatory diseases, pulmonary diseases and cardiovascular diseases.