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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

In stock

$415.00

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.
Garg, Manisha et al. (2017) Liraglutide acutely suppresses glucagon, lipolysis and ketogenesis in type 1 diabetes, Diabetes Obes Metab. 2017 Mar 17.
Goldgof et al., The Chemical Uncoupler 2,4-Dinitrophenol (DNP) Protects against Diet-induced Obesity and Improves Energy Homeostasis in Mice at Thermoneutrality. J. Biol. Chem., Jul 2014; 289: 19341 - 19350.
Ji et al., Molecular and metabolic profiles suggest that increased lipid catabolism in adipose tissue contributes to leanness in domestic chickens.Physiol Genomics, May 2014; 46: 315 - 327.
Takimoto & Hamada. Acute exercise increases brain region-specific expression of MCT1, MCT2, MCT4, GLUT1, and COX IV proteins. J Appl Physiol, May 2014; 116: 1238 - 1250.
Anja Solberg et al., Deletion of mouse Alkbh7 leads to obesity. J Mol Cell Biol, Jun 2013; 5: 194 - 203.
For more citations of this product click here
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