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beta Hydroxybutyrate (beta HB) Assay Kit (Colorimetric) (ab83390)

Submit a review Q&A (17)References (32)
Functional Studies - beta Hydrobutyrate (beta HB) Assay Kit (ab83390)
  • Functional Studies - beta Hydrobutyrate (beta HB) Assay Kit (ab83390)
  • Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)
  • Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)
  • Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 40 min
  • Sample type: Cell culture supernatant, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
  • Sensitivity: 0.01 mM

Overview

  • Product name

    beta Hydroxybutyrate (beta HB) Assay Kit (Colorimetric)
    See all Beta Hydroxybutyrate kits

  • Detection method

    Colorimetric

  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts

  • Assay type

    Quantitative

  • Sensitivity

    > 0.01 mM

  • Range

    0.01 mM - 0.2 mM

  • Assay time

    0h 40m

  • Product overview

    beta Hydroxybutyrate (beta HB) Assay Kit (Colorimetric) (ab83390) utilizes beta HB Dehydrogenase to generate a product which reacts with a colorimetric probe with an absorbance band at 450 nm.


    The kit is an easy and accurate assay to measure beta HB levels in biological samples. The assay is linear for 1-20 nmol beta HB in up to 100 µL samples or 0.01-0.2 mM of beta HB samples.


    beta Hydroxybutyrate assay protocol summary:
    - add reaction mix to sample and standard wells
    - incubate for 30 min
    - analyze with microplate reader

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K632 β-Hydroxybutyrate (β-HB) Colorimetric Assay Kit. K632-100 is the same size as the 100 test size of ab83390.

    During diabetic ketoacidosis the liver produces ketone bodies (acetoacetate, acetone, and primarily beta-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.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate reader

Properties

Associated products

Images

  • Functional Studies - beta Hydrobutyrate (beta HB) Assay Kit (ab83390)
    Functional Studies - beta Hydrobutyrate (beta HB) Assay Kit (ab83390)Image from Zhao M et al., PLoS One. 2017;12(3):e0174477. Fig 1(E); doi: 10.1371/journal.pone.0174477. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    The plasma levels of total BHB was detected using ab83390. Five rats were in each group. Samples were collected from tail vein blood and were transferred into EDTA-containing tubes. After centrifugation at 1500×g for 15 min at room temperature, the plasma was transferred into a new tube and stored at -80°C until use. Values are presented as the mean ± S.E. (**p<0.01).

  • Functional Studies - beta Hydrobutyrate (beta HB) Assay Kit (ab83390)
    Functional Studies - beta Hydrobutyrate (beta HB) Assay Kit (ab83390)Image from Wakade C et al., PLoS One. 2014;9(10):e109818. Fig 2(D).; doi: 10.1371/journal.pone.0109818. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Human serum samples were filtered using a 10 kda MW cut-off spin filter (ab93349) spun at 14,000 RPM for 30 minutes. Beta Hydrobutyrate (BHB) was detected on a 96-well plate using ab83390.

    Young control group (6 men, 25±1 years old).
    Age-matched control group (13 men, 69±7 years old)
    PD = Individuals diagnosed with idiopathic PD (15 men, 71±8 years old, duration of disease = 10±6 years, H&Y = 2.3±0.7, median = 2).

    A trend approaching significance in increased levels of BHB in the plasma of PD patients compared to age-matched controls was observed. BHB is known to fluctuate in a variety of conditions including starvation, diabetes mellitus and other co-morbidities.

  • Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)
    Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)

    Beta hydroxybutyrate in mouse tissue lysates showing quantity (nmol) per mg protein of tested sample

  • Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)
    Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)

    Beta hydroxybutyrate measured in biological fluids showing quantity (nmol) per microliter of tested sample

  • Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)
    Functional Studies - beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)
    beta HB Standard Curve: Assays were performed following ab83390 kit protocol.

References (32)

Publishing research using ab83390? Please let us know so that we can cite the reference in this datasheet.

ab83390 has been referenced in 32 publications.

  • Perrotta P  et al. PFKFB3 gene deletion in endothelial cells inhibits intraplaque angiogenesis and lesion formation in a murine model of venous bypass grafting. Angiogenesis N/A:N/A (2021). PubMed: 34432198
  • Perrotta P  et al. Partial Inhibition of Glycolysis Reduces Atherogenesis Independent of Intraplaque Neovascularization in Mice. Arterioscler Thromb Vasc Biol 40:1168-1181 (2020). PubMed: 32188275
  • Bocco BMLC  et al. Iodine Deficiency Increases Fat Contribution to Energy Expenditure in Male Mice. Endocrinology 161:N/A (2020). PubMed: 33091112
  • Hasenour CM  et al. Vitamin E does not prevent Western diet-induced NASH progression and increases metabolic flux dysregulation in mice. J Lipid Res 61:707-721 (2020). PubMed: 32086244
  • Shippy DC  et al. ß-Hydroxybutyrate inhibits inflammasome activation to attenuate Alzheimer's disease pathology. J Neuroinflammation 17:280 (2020). PubMed: 32958021
View all Publications for this product

Customer reviews and Q&As

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1-10 of 17 Abreviews or Q&A

Question

I would like to test beta hydroxybutyrate and am quite insterested in your kit (ab83390). From the protocol, I found dehydrogenase was used to oxidize hydroxybutyrate first and then a detection reagent will recognize oxobutyrate.

Here, I am wondering whether the detection is specific for oxobutyrate. If there are any compounds in the mixture contains ketone, will they be detected also?

Read More
Answer

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 hydroxybutyrate and will not recognize other ketones.

Read More

Question

Hello. Can you tell me if the beta HB assay will also measure HB esters such as methyl 3 hydroxy butyrate or butyrate dimers?

Thank you,

Read More
Answer

Thank you for your enquiry and interest in our products.

I can confirm that this kit (ab83390: beta Hydroxybutyrate (beta HB) Assay Kit) measuresonly the β-Hydroxybutyrate in the samples.

Hope this information has been helpful for you.

Read More

Question

High background using this kit with fish plasma samples.

Read More
Answer

Thank you for your phone call a couple of weeks ago, and I sincerely apologize for the delay getting back to you.

I've forwarded your data to our assay expert in the lab, and the kit has been standardized using mammalian samples so it may require some additional optimization to be used with fish samples. To reduce the background, we would recommend using the plasma undiluted in at 50-100 uL per well. Assay buffer would not be added to further dilute the samples. We would also recommend filtering the plasma with a 10 kDa molecular weight cutoff filter to help remove interfering substances. We have ab93349 which can be used for this purpose-

https://www.abcam.com/10kD-Spin-Column-ab93349.html

We do fully guarantee the results of this kit, so I would be happy to send a replacement or issue a credit or refund if the results are not satisfactory.

I hope that these suggestions will be helpful, but please let me know if you have further questions or if there is anything else that we can do for you, and I'll be happy to help.

Read More

Question

I have order two Abcam kits to measure beta-HB and Ammonia in mouse serum sample--beta Hydroxybutyrate (beta HB) Assay Kit (ab83390) and Ammonia Assay Kit (ab83360). Do you have any special suggestions on how to prepare blood sample?

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Answer

For serum or plasma samples, there are no special requirements for preparation of serum or plasma samples for the ammonia assay. For the beta HB assay, it is recommended (see section 4 of the protocol) that samples be de-proteinated with a 10 kDa filter, which we have as item ab93349.

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Question

using the ab93349 assay kit to detect hydroxybutyrate. I understand that the enzyme provided in the kit is the hydroxybutyrate dehydrogenase. In order for the kit to work, is the assumption made that the biological fluid being analyzed has the diaphorase that oxidizes the NADH and transfers electrons to the detection dye? We are simply trying to better understand the principle of the technique and how to make sure we are including appropriate controls.

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Answer

The principle you have correct, but the samples need not have any diaphorase. Everything required for this assay to work is being provided in the kit components.

I hope this information has been useful for you. Please let me know if you have any other questions.

Read More

Question

Hello

I'd like to confirm that your
beta Hydroxybutyrate (beta HB) Assay Kit (ab83390)
can be used with plasma (serum) samples that were frozen and stored at -80C. The samples have not undergone any freeze-thaw cycles. Please let me know if frozen samples are OK or whether they must be fresh samples for best results.

Thank you,
--

Read More
Answer

Thank you for your inquiry.
I can confirm that frozen serum or plasma samples are suitable for this kit.
We did not experimentally test frozen versus fresh samples but would expect a slight loss of activity in frozen samples.
This will not impact on your results if you are only comparing frozen samples.
I hope this information is helpful and wish you good luck with your research.

Read More

Question

Dear Sir/madam,

I am interested the kit you provides.

We plane to measure the beta Hydroxybutyrate from tissue.

In the instruction saying it can be used to measure tissue extract.

Can I ask you, what kind of the buffer for tissue extract? Do I need to purchase spin column for this purpose?

Thank you

Best wishes

Read More
Answer

Thank you for your enquiry.

I can confirm that tissue samples in the form of homogonate can be used with this kit.

In this case, carefully cut up the tissue to small pieces, then lyse using the kit Assay Buffer. To remove interfering substances, the sample can be spun filtered with a 10 kDa molecular weight cut off spin filter.

ab93349 10kD Spin Column
https://www.abcam.com/10kD-Spin-Column-ab93349.html (or use the following:
https://www.abcam.com/10kD-Spin-Column-ab93349.html).

For unknown samples, it may be necessary to test several different doses to ensure the readings are within the range of the standard curve. You may also need to correct the background contributions from NADH or NADPH in the sample. This is done as follows:

'Reduced pyridine nucleotides NAD(P)H can interfere with the assay. If the presence of these compounds is suspected in the sample, run a background control substituting the 2 ul Enzyme Mix with 2 ul Assay Buffer. The background reading should be subtracted from β-HB sample reading.'

I hope this will be helpful to you. Should you have any further questions, please do not hestitate to contact us.

Read More

Question

- Can we use XXXX 24kDa cut off filters?
- In assay protcol step 4 the readings increases after incubation 30 minutes and rises untill 7 hours.
- Sample used were Drosophila homogenates
- All the reading were in standard range

Read More
Answer

Thank you for contacting us.

You can certainly use the spin column from XXXXXX however the procedure should be followed as per manufacturer's datasheet.

Regarding the high readings, this assay has not been standardized for drosophila samples. So 30 min might not be the right incubation time. The times have to be standardized. Another thing to keep in mind is that the colour is probably overdeveloping, as long as the colour is within the detection range of the machine and linear range of the standard curve, it should be fine.

We may suggest you to take readings at different time points,for getting more data from this kit.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

Inquiry: We are interested in purchasing one assay kit for beta hydroxybutyrate (ab83390). Please give us instructions to place the order from Spain Thank you in advance

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Answer

Gracias por contactarnos.

Puede contactarnos directamente en español, telefónicamente (91 114 65 54), o bien por correo electrónico, a:

mailto:orders@abcam.com Para tramitar pedidos y otras consultas relacionadas con Servicio al Cliente

mailto:soportecientifico@abcam.com Para consultas y reclamaciones científicas.

Para realizar un pedido puede hacerse por teléfono, fax (91 114 65 55), email o a través de la website.

Es importante indicar la siguiente información:

- dirección completa (para el envío y para la facturación)

- número de teléfono y de fax, además de una dirección de e-mail

- nombre de la persona a la cual debemos enviarle los productos

- su número de identificación fiscal (NIF)

- número de catálogo del producto y su descripción correspondiente

Si tenéis alguna otra consulta, no dudéis en contactarnos de nuevo.

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Question

can rat, mouse and monkey samples be used with this kit?

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Answer

Thank you for contacting us.

I have confirmed with the lab thatthis kit is indeed compatible with all mammalian samples. You can definitely use it with rat, mouse and monkey samples.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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1-10 of 17 Abreviews or Q&A

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