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

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.

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.

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.

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.

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.

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.

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.

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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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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