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β-Hydroxybutyrate (β-HB) Colorimetric Assay Kit

based on 13 citations in multiple journalsβ-Hydroxybutyrate (β-HB) Colorimetric Assay Kit135 5
Catalog #: K632
$450.00

Product Details

Cat # +Size K632-100
Size 100 assays
Detection Method Absorbance (450 nm)
Species Reactivity Mammalian
Applications The assay is linear for 1-20 nmol β-HB in up to 100 µl samples or 0.01-0.2 mM of β-HB samples.
Features & Benefits • Simple procedure; takes ~ 40 minutes
• Fast and convenient
• The kit provides an accurate assay for measuring β-Hydroxybutyrate (β-HB) in various sample
Kit Components • β-HB Assay Buffer
• β-HB Enzyme Mix
• β-HB Substrate Mix
• β-HB Standard (1.0 µmol)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Diabetic ketoacidosis occurs when circulating insulin levels drop to very low levels, shutting off the supply of glucose to the body. The physiological response is for the liver to produce ketone bodies (acetoacetate, acetone, and primarily β-hydroxybutyrate) from the acetyl CoA produced from fatty acid oxidation. The very high rate of ketone body production outstrips the body’s ability to utilize them as an energy source and the blood concentration builds up. As rather strong acids, they result in a significant drop in blood pH. BioVision’s β-HB Assay kit utilizes β-HB Dehydrogenase to generate a product which reacts with our colorimetric probe with an absorbance band at 450 nm. The kit is an easy and accurate assay to measure β-HB levels in biological samples. The assay is linear for 1-20 nmol β-HB in up to 100 µl samples or 0.01-0.2 mM of β-HB samples.


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Which anticoagulant is recommended for blood collection for this assay?
Heparin can be used.
How should plasma be processed for thi s assay?
Plasma sample can be used directly or filtered depending upon the amount of insoluble cloudy components.
I found dehydrogenase was used to oxidize hydroxybutyrate first and then a detection reagent will recognize oxobutyrate.I am wondering whether the detection is specific for oxobutyrate. If there are any ketones, will they be detected also?
The detailed assay chemistry is proprietary information. 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.
Should urine samples be filtered before use?
We recommend filtering the urine to remove any particulate matter.
How are cells/tissues prepared for this assay?
Start with 10-100 mg of the tissue, add 500-1000 µl (or ~4-6 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (30-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample at 4C for 15 mins at 10,000 RPM and collect the supernatant. Load the supernatant unto a 10kda spin column for deproteinzation. Use the eluate for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.
What cell lysate volume should be used for the assay?
The volume used is sample dependent and has to be optimized by the user. We recommend that you run a pilot assay using different volumes of the samples such that the readings are within the linear range of the standard curves
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted
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 the appropriate temperature and appropriate conditions. 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 alternate buffers be used for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
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.
Can I use K808 for K632-100 beta Hydroxybutyrate (beta HB) Assay Kit (Colorimetric) assay sample deproteinization?
No, we recommend using the 10 kDa spin filters (BV Cat# 1997) to deproteinize the samples for K632.
I have limited tissue samples and would like to use same samples for K627-100 (L-Lactate Assay Kit (Colorimetric)) and K632-100 beta (Hydroxybutyrate (beta HB) Assay Kit (Colorimetric)). The protocols for the two kits are similar. Could you please confirm if the assay buffers provided in those two kits are the same?
Unfortunately, the assay buffers are not the same. We recommend using the assay buffers that are included with each kit to be used for preparing samples for that assay.
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.
Zhou Zhou, Adipose‐Specific Lipin‐1 Overexpression Renders Hepatic Ferroptosis and Exacerbates Alcoholic Steatohepatitis in Mice. Hepatol Commun, May 2019; 31061954.
Cuiying Xiao, Physiology and effects of nucleosides in mice lacking all four adenosine receptors. PLoS Biol., March 2019; 30822301.
C Joseph Burnett, Need-based prioritization of behavior. eLife, March 2019;  30907726.
Frida A. Teran, Time of Day and a Ketogenic Diet Influence Susceptibility to SUDEP in Scn1aR1407X/+ Mice. Front Neurol., March 2019; 30984098.
Yayun Liang, et al., (2018) A combination of p53-activating APR-246 and phosphatidylserine-targeting antibody potently inhibits tumor development in hormone-dependent mutant p53-expressing breast cancer xenografts, Dove Med Press, Mar. 2018, 29606888
For more citations of this product click here