BHB-Glo™ (Ketone Body) Assay
Easy, Sensitive Beta-Hydroxybutyrate Assay Kit
- Compatible with cells, media, tissue and serum samples
- In-well sample preparation protocol streamlines workflow
- Scalable to 384-well plates for HTS applications
Catalog Number:
Size
Catalog Number: JE9500
Catalog Number: JE9600
Efficient Detection of BHB in Diverse Samples
The BHB-Glo™ (Ketone Body) Assay is a fast and sensitive method for detecting β-hydroxybutyrate (BHB) from various biological samples, such as cultured cells, media, tissue homogenates and serum. Based on bioluminescent technology, the BHB-Glo™ Assay can detect subtle changes in ketosis as well as ketone body consumption and secretion. Its streamlined sample preparation protocol involves acid treatment and neutralization directly in the assay wells, eliminating the need for cell collection, centrifugation, spin columns and deproteinization. The assay is adaptable to a 384-well format, facilitating quick analysis of many samples.
How the BHB-Glo™ Assay Works
β-Hydroxybutyrate (BHB) Dehydrogenase catalyzes the oxidation of BHB, simultaneously reducing NAD+ to NADH. In the presence of NADH, Reductase converts the proluciferin Reductase Substrate into luciferin. This reaction is detected by Ultra-Glo™ Recombinant Luciferase, which emits light proportionate to the amount of BHB in the sample.
Simple Add-and-Read Protocol Saves Time
Accurately Detect the Smallest Changes in BHB
|
Sensitivity (S/B >5) |
800nM |
|
|
Limit of Detection (S/N >3) |
50nM |
|
|
Linear Range |
50nM to 25μM |
|
|
Maximum Assay Window (S/B) |
>200 |
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Measure BHB Secretion from Hepatocytes
The BHB-Glo™ Assay can be used to monitor intracellular and extracellular BHB levels in cell culture. In the data shown, HepG2 cells were plated in a 96-well plate, and then media samples were removed at prescribed time points to monitor BHB secretion over time. Extracellular BHB was measured using the BHB-Glo™ Assay. Greater than sixfold signal above background was observed at 24 hours.
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Protocols
Complete Protocol
Specifications
Catalog Number:
What's in the box
| Item | Part # | Choose a size | Concentration |
|---|---|---|---|
β-Hydroxybutyrate Dehydrogenase |
J414A | 1 × 100μl | |
β-Hydroxybutyrate, 10mM |
J415A | 1 × 50μl | |
Reductase |
G884A | 1 × 55μl | 6mg/ml |
Reductase Substrate |
G885A | 1 × 55μl | |
Luciferin Detection Solution |
J135A | 1 × 5ml | |
NAD |
J136B | 1 × 275μl | |
Neutralization Buffer |
J153A | 1 × 15ml | |
NADP+ |
J154B | 1 × 100μl | |
0.6N HCl |
J402A | 1 × 15ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
What's in the box
| Item | Part # | Choose a size | Concentration |
|---|---|---|---|
β-Hydroxybutyrate Dehydrogenase |
J414B | 1 × 1ml | |
β-Hydroxybutyrate, 10mM |
J415A | 1 × 50μl | |
Reductase |
G884B | 1 × 275μl | 6mg/ml |
Reductase Substrate |
G885B | 1 × 275μl | |
Luciferin Detection Solution |
J135B | 1 × 50ml | |
NAD |
J136C | 1 × 1ml | |
Neutralization Buffer |
J153A | 1 × 15ml | |
NADP+ |
J154C | 1 × 500μl | |
0.6N HCl |
J402A | 1 × 15ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
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