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PicoProbe™ β-Hydroxybutyrate Fluorometric Assay Kit

The most sensitive kit on the market
Catalog #: K651

Product Details

Cat # +Size K651-100
Size 100 assays
Detection Method Fluorescence (Ex/Em = 535/587 nm)
Applications Measurement of β-Hydroxybutyrate in various biological samples
Analysis of β-Hydroxybutyrate in pathological conditions
Features & Benefits • Simple, rapid & high-throughput adaptable
• Highly sensitive: The assay can detect less than 4 µM of β-Hydroxybutyrate (β-HB) in various sample types.
Kit Components • β-HB Assay Buffer
• PicoProbe™ (in DMSO)
• β-HB Enzyme Mix (Lyophilized)
• β-HB Substrate Mix (Lyophilized)
• β-HB Standard (Lyophilized)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


β-Hydroxybutyrate (β-HB) is the most abundant of the ketone bodies (~78% of total ketone bodies in blood). Ketoacidosis (high concentrations of the ketone bodies) can be caused by a variety of conditions, such as diabetes, alcoholism and severe starvation. Ketoacidosis can be fatal if not treated. Additionally, ketogenic diets can increase the concentration of ketone bodies slightly in healthy individuals. BioVision’s β-Hydroxybutyrate Assay Kit offers simplicity, enhanced sensitivity, and can be adapted to high-throughput applications. The assay is based on enzymatic oxidation of β-HB that results in generation of fluorescent signal (Ex/Em = 535/587 nm) which is directly proportional to the amount of β-HB. The kit offers an excellent alternative for measurement of β-HB when biological sample quantities are limited or subketogenic levels are suspected. The assay can detect β-Hydroxybutyrate as low as 4 µM in a variety of biological samples.

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What is the speciality of this kit versus K632?
This kit can detect less than 4 µM of ß-Hydroxybutyrate (ß-HB) and is the most sensitive kit on the market. It uses the proprietary Picoprobe and uses fluorometric detection. K632 detects 0.01-0.2 mM of ß-HB and is a colorimetric method.
Does this kit detect oxobutyrate or other ketones?
The detection agent does not quantify the oxobutyrate but the byproduct of the reaction by the dehydrogenase. The reaction byproducts are produced proportionally based on the amount of beta hydroxybutyrate in a specific sample. This is why this assay kit is specific for beta hydroxy butyrate and will not recognize other ketones.
Which anticoagulant is recommended for blood collection for this assay?
Heparin can be used.
Can plasma/serum samples be stored for doing the assay later?
Although not ideal , plasma/serum can be stored at -80C until the assayis performed. We did not experimentally test frozen versus fresh samples but would expect a slight loss of activity in frozen samples.This should not impact the results when comparing frozen samples.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at -20C throughout. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Have you tested adherent cells with this kit? If so, do you have any protocol recommendations?
We have not tested adherent cells with this kit. Theoretically you should be able to use adherent cells with the kit. For sample preparation, you can start with 1x10^6 cells and homogenize in 100 µl of cold B-HB assay buffer. Homogenize using either a dounce homogenizer or sonicate for few short pulses on ice. Centrifuge at 10,000g for 10 minutes and take the supernatant. Filter the supernatant through a 10 kDa spin filter and use that for the assay.