protein carbonyl in specimen (OxyBlot assay)

Procedure

• carbonyl groups on the protein side chains are derivatized to 2,4-dinitrophenylhydrazone (DNP-hydrazone) by reaction with 2,4-dintrophenylhydrazine
• the DNPH-derivatized proteins are separated by polyacrylamide gel electrophoresis (PAGE) followed by Western blotting
• alternatively, a dot-blot method can be used
• filter blots are then incubated with primary antibody (rabbit IgG) to DNP followed by incubation with a peroxidase-conjugated goat anti-rabbit IgG
• filters are then treated with chemiluminescent reagents (luminol & enhancer)
• light emission at 428 nm is detected on blue light-sensitive film after addition of hydrogen peroxide

More general terms

• special chemistry test

More specific terms

• protein carbonyl in serum/plasma

Additional terms

• protein carbonyl