Description
beta-Hydroxybutyrate Dehydrogenase (BDH) Activity Assay Kit
The Assay Genie beta-Hydroxybutyrate Dehydrogenase (BDH) Activity Assay Kit measures BDH (3-hydroxybutyrate dehydrogenase, BDH1, EC 1.1.1.30) activity in cell and tissue lysates and serum. This mitochondrial enzyme interconverts acetoacetate and β-hydroxybutyrate, the two main ketone bodies, and is central to ketogenesis in the liver and ketone body utilisation in heart, brain and muscle.
NADH produced by BDH drives the reduction of the tetrazolium dye INT to a cherry-red formazan, measured at 492 nm in a 96-well plate. Each sample is run alongside a no-substrate control, and activity is calculated directly in IU/L, making the kit a convenient tool for studies of fasting, ketogenic diets, diabetes and cardiac metabolism.
| Product Name: | beta-Hydroxybutyrate Dehydrogenase Assay Kit |
| SKU: | BR00004 |
| Kit Size: | 100 wells |
| Detection Method: | Colorimetric (INT → formazan, OD 492 nm) |
| Sample Types: | Cell and tissue lysates, serum (diluted 10x); not for plasma |
| Recommended Protein: | 0.5–3 mg/ml |
| Incubation Time: | 60 or 120 min at 37 °C |
| Units: | IU/L (µmol/L/min); normalise to protein or GAPDH |
| Equipment Required: | Microplate reader (492 nm) |
| Storage: | -80 °C (see component storage) |
| Intended Use: | For research use only (RUO). Not for diagnostic procedures. |
- Direct activity calculation in IU/L, no standard curve required
- Compatible with lysates and diluted serum
- Per-sample no-substrate control removes background
- Simple mix-and-read 96-well format
- Includes 10x Cell Lysis Solution
- BDH oxidises β-hydroxybutyrate to acetoacetate, reducing NAD+ to NADH.
- NADH drives the reduction of INT to a coloured formazan. Formazan has an absorbance maximum at 492 nm (ε = 18 mM-1cm-1), so activity is calculated directly in IU/L (µmol/L/min) without a standard curve.
- Activity is calculated from ΔOD (reaction − control): IU/L = ΔOD × 8.1 (60 min) or × 4.05 (120 min).
- Cells: ~105 washed cells in 50–100 µl ice-cold 1x Cell Lysis Solution, 5 min on ice.
- Tissue: ~10 mg tissue homogenised in 0.5 ml ice-cold 1x Cell Lysis Solution.
- Centrifuge at ~14,000 rpm for 5 min; adjust protein to 0.5–3 mg/ml.
- Serum: dilute 10-fold with ice-cold dH2O. Do not use plasma.
| Component | Size | Storage |
| BDH Assay Solution | 5 ml | -80 °C, protect from light |
| 10x BDH Substrate | 0.5 ml | -20 °C (short term) or -80 °C |
| 10x Cell Lysis Solution | 25 ml | Room temperature |
- Ketone body metabolism, ketogenesis and ketolysis
- Fasting, caloric restriction and ketogenic diet studies
- Diabetes and metabolic disease research
- Cardiac and brain energy metabolism
Can I use plasma with the BDH Activity Assay Kit?
No. The manual recommends cell/tissue lysates or serum diluted 10-fold; plasma should not be used.
How is BDH activity calculated?
Subtract the control well from the reaction well to obtain ΔOD at 492 nm, then multiply by 8.1 (60 min) or 4.05 (120 min) to get IU/L.
What equipment do I need?
A 492 nm microplate reader, a 37 °C (non-CO2) incubator, a refrigerated microcentrifuge and a plain 96-well plate.
