QuantiChrom™ Carbonyl Assay Kit
Protocol
SDS
Application
- Quantification of carbonyl groups or protein carbonyls in biological samples.
Key Features
- Fast and sensitive. Use of 100 µL sample. Linear detection range from 12 to 250 µM carbonyl in 96-well plate assay.
- Convenient. The procedure involves adding a single working reagent, and reading the absorbance after 30 min.
- High-throughput. Homogeneous "mix-incubate-measure" type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.
Method
- OD375nm
Samples
- Chemicals with carbonyl groups (e.g. ketones, aldehydes) or protein carbonyls in biological samples (e.g. oxidized BSA, etc.)
Species
- All
Size
- 100 tests
Detection Limit
- 12 µM
Shelf Life
- 12 months
More Details
- CARBONYL groups, such as ketones and aldehydes, are a commonindicator of protein oxidation. Protein oxidation is caused as a result of exposure to reactive oxygen species (ROS) and is a common marker in various diseases as well as aging. BioAssay Systems' Carbonyl Assay Kit is based on an improved method, where 2,4-dinitrophenylhydrazine (DNPH) reacts with carbonyl groups to produce a colored compound at 375 nm. The intensity of this colored compound is directly proportional to the carbonyl groups in the sample.
The background is very high. Is this normal?
The background absorbance is typically ~ 0.5. This is completely normal.Can I store the diluted standards (250 µM, 150 µM, 75 µM, and 0 µM) to use for later?
We recommend preparing the standards fresh from the 50 mM each day.Do I need to run a standard curve with every assay?
Yes.How should I store the kit?
The kit is shipped at room temperature. Upon delivery, store at -20°C.Precipitate formed when I added the reagent to the sample. What should I do?
The Reagent is acidic and may cause precipitation in samples that have significant amounts of protein. You may need to dilute your sample, repeat the assay, and correct for the dilution factor in the calculation.What is the Carbonyl Standard?
The standard is pyruvate, which has two carbonyl groups per molecule. So the 50 mM Carbonyl Standard is actually 25 mM pyruvate.My sample has high levels of nucleic acids. Will this interfere?
Nucleic acids can artificially increase the carbonyl count if they are present in high levels. If the ratio of 280/260 nm is less than 1, samples should be mixed with 1% streptomycin sulfate to denature the nucleic acids. Mix well, incubate the samples at room temperature for 15 minutes and then centrifuge at 5000xg for 10 minutes. Use the supernatant in the assay.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.
Pena-Bermudez, Y. A., et al. (2020). Effects of feeding increasing levels of yerba mate on lamb meat quality and antioxidant activity. Animals: An Open Access Journal from MDPI 10(9). Assay: Carbonyl Groups in lamb longissimus thoracis muscle.
To find more recent publications, please click here.
To find more recent publications, please click here.
If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.
- Fast turnaround
- Quality data
- Low cost
Please email or call 1-510-782-9988 x 2 to request assay service.
- Fast turnaround
- Quality data
- Low cost
Please email or call 1-510-782-9988 x 2 to request assay service.