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

    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.

  • Platform
    Microplate reader

Properties

Associated products

Images

  • 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).

  • 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.

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

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

  • beta HB Standard Curve: Assays were performed following ab83390 kit protocol.

Protocols

References

This product has been referenced in:
  • Liu SJ  et al. FTO is a transcriptional repressor to auto-regulate its own gene and potentially associated with homeostasis of body weight. J Mol Cell Biol N/A:N/A (2018). Read more (PubMed: 29771336) »
  • Tumurkhuu G  et al. Chlamydia pneumoniae Hijacks a Host Autoregulatory IL-1ß Loop to Drive Foam Cell Formation and Accelerate Atherosclerosis. Cell Metab 28:432-448.e4 (2018). Read more (PubMed: 29937375) »
See all 11 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

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.

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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.

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Question
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.

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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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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.

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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.

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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/index.html?datasheet=93349 (or use the following:
https://www.abcam.com/index.html?datasheet=93349).

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.

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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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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
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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