Overview

  • Product name

    Aldehyde Assay Kit (Colorimetric)
    See all Aldehyde kits
  • Detection method

    Colorimetric
  • Sample type

    Saliva, Plasma, Cell culture extracts, Adherent cells, Suspension cells
  • Assay type

    Quantitative
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Aldehyde Assay Kit (Colorimetric) ab112113 is a a sensitive, one-step colorimetric assay to detect as little as 1 nanomole of aldehyde in a 100 µL assay volume (10 µM). The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and readily adapted to automation without a separation step.


    The aldehyde assay protocol uses a proprietary dye that generates a chromogenic product upon reacting with an aldehyde. Most of the existing aldehyde test methods are based on separations either by the tedious and expensive HPLC-MS or GC-MS.


    The signal is read with an absorbance microplate reader at 405 or 550 nm.


    Aldehyde assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 30-60 min
    - analyze with a microplate reader

  • Notes

    Previously called Aldehyde Quantification Assay Kit (Colorimetric).

    This kit has been used for monitoring activities of oxidases that convert an amino group to an aldehyde group.

    ab112113 should be stored desiccated.

  • Platform

    Microplate reader

Properties

Images

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Aldehyde measured in biological fluids showing quantity (uM) per mL of tested sample. Values calculated based on the 550 nm Standard Curve. Samples diluted 1-10 fold.

  • Aldehyde measured in cell culture supernatants showing quantity (uM) per mL of tested sample. Values calculated based on the 405 nm Standard Curve. Samples diluted 2 fold.

  • Aldehyde measured in cell lysates showing quantity (uM) per 1 mln cells. Values calculated based on the 550 nm Standard Curve.

    Samples with the concentration of 1.26e8 cells/mL (HeLa) and 1e8 cells/mL (HepG2) were used. Samples were diluted 3 fold.

  • Aldehyde dose response was measured in a 96-well black plate with ab112113. As low as 10 µM (1 nanomol/well) of aldehyde can be detected with 30 minutes incubation (n=3). Standard curve was read at 405 nm.

Protocols

References

This product has been referenced in:

  • Li C  et al. Spermine synthase deficiency causes lysosomal dysfunction and oxidative stress in models of Snyder-Robinson syndrome. Nat Commun 8:1257 (2017). Read more (PubMed: 29097652) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 22 Abreviews or Q&A

Question
Answer

Sugar should be OK, but thiols will interfere with the assay.

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Answer

ab112113 detects all the aldehydes in solution. We don't have experiences in tissue samples, but it should be ok theoretically. Simply use your traditional homogenization method for sample preparation.

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Answer

For product ab112113, you measure the endpoint and not kinetic change over time. After 30-60 minutes of incubation this is when you can measure the OD at 405 or 550 nm depending on on the concentration of aldehyde in your samples. At lower concentrations, the absorbance at 405 nm gives better results. We would recommend to therefore read at 405nm and this should be fine but if you have high concentrations of aldehyde, you can read at 550 nm as this may give a better reading.

In addition, you can do a time course for the initial measurement to see which time point gives the best result. In general an OD at around ˜ 0.2-0.5 should be the best time for your measurement.



This kit should indeed measure all known aldehydes including the detection of polysaccharide aldehydes.

For saliva samples there is no special pre-treatment and they can simply be treated like the standards.

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Answer

The assay ab112113 is capable of detecting aldehydes in polysacharrides but we have not tested that sample type. The standard in the assay is glyceraldehyde which is relatively small, because the assay was developed to detect small aldehydes. So, detection of polysaccharide aldehydes is expected, but we do not know how the signal that develops will compare to the signal from the standards.

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Answer

Thank you for contacting us.



1. The wavelength is mostly depending on the type of the aldehyde, and the incubation time. For example, Formaldehyde have only one peak 550nm, others have two peaks.

2. Our standard is

800x600

Glyceradehyde, your sample's aldehyde might be different from the standard, that is probably the reason why the color is different (such as FBS has more 405 compare to 550nm). There are 2 products after reacted with the dye, 405nm at beginning (yellow), and then it tends to 550 nm (brownish-red) after longer incubation time such as 1 hour. So use 550 nm for both standard and your samples should give you more accurate results.

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Answer

The lab has not observed this before, and we have not received similar feedback from other customers. I would choose the values you obtained for the shorter assay incubation, rather than the overnight one. What you are seeing over time may in fact be the production of acetaldehyde in the assay wells over time, but we do not have an explanation for how this might happen.


My reason for choosing the values from the shorter incubation (30-60 minutes of incubation is what is suggested in the protocol) is that they are more likely to reflect the amounts present in the cells at the time of harvest, more so than the values from the later (overnight incubation) timepoint, which seems to include signal produced by an assay-induced or lysis-induced process. Have you considered assaying samples that were held overnight at the assay temperature, before assaying them? Comparison of these sample with samples assayed immediately after harvest might reveal production of acetaldehyde, independent of the assay reagents.

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Answer

ab112113 can be used on plasma samples. The plasma samples don't need any special preparation so no spin columns are needed to deproteinise these samples.
.

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Answer

The lab does not have experiences using this on fermentation broth. It should be OK theoretically, but you will need to subtract the background color of the broth from your reads.

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Answer

Hello and thank you for contacting us. The kit should work using the method you described, stopping the reaction, but we recommend using a basic solution (pH8) when doing so. Please let me know if you have any further questions.

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Answer

Thank you for your inquiry.
I can confirm that the standard of ab112113 is Glyceradehyde (MW=90).
I hope this information is helpful and that this kit is suitable for your experiments.

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1-10 of 22 Abreviews or Q&A

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