EnzyChrom™ α-Ketoglutarate Assay Kit
Application
- For quantitative determination of αKG in serum, tissue and cell lysates.
Key Features
- Sensitive and accurate. Linear detection range in 96-well plate: 1.3 to 200 μM for colorimetric assays and 0.08 to 20 μM for fluorimetric assays.
- Fast and convenient. Room temperature assay with a 30 min incubation. No 37°C incubator is needed.
- High-throughput. Homogeneous “mix-incubate-measure” type assay. Can be readily automated to assay thousands of samples per day.
Method
- OD570nm or FL530/585nm
Samples
- Serum, tissue and cell lysate
Species
- All
Procedures
- 30 min
Size
- 100 tests in 96-well plate
Detection Limit
- 1.3 μM for OD, 0.08 μM for FL
Shelf Life
- 6 months
More Details
α-Ketoglutarate (αKG) is a crucial intermediate in the citric acid (TCA) cycle and plays an important role in amino acid formation, nitrogen transport, and oxidation reactions. αKG can modulate muscle and bone growth as well as the aging process, and has also been investigated for its potential to enhance anticancer immune functions. BioAssay Systems’ α-ketoglutarate assay uses a single Working Reagent that combines αKG deamination, pyruvate oxidation and hydrogen peroxide determination. The change in OD570nm or fluorescence intensity at λex/em = 530/585nm is directly proportional to the αKG present in the sample.Can linear curve fitting be used?
While linear curve fitting can be applied in theory, we recommend using the internal standard method as described in the protocol. This approach accounts for sample matrix effects and improves accuracy and reproducibility, especially when working with complex biological samples.
Do urine samples work with this assay?
Urine samples are confirmed to be incompatible with this assay, possibly due to high salt and interfering metabolites in urine, including pyruvate.
When tissue is used, should it be freshly obtained? Or is -80°C storage ok?
If direct sample processing is not possible, we recommend snap freezing tissue samples in liquid nitrogen and keeping them either in liquid nitrogen or at -80°C until further processing.
For more detailed product information and questions, please feel free to email or call us at 1-510-782-9988 x 2. For more general information regarding our assays, please refer to Technical Support.
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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
