Overview

  • Product name
    Urea Assay Kit
    See all Urea kits
  • Detection method
    Colorimetric
  • Sample type
    Urine, Serum, Plasma, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay type
    Quantitative
  • Sensitivity
    > 0.5 nmol/well
  • Range
    0.5 nmol/well - 5 nmol/well
  • Assay time
    1h 00m
  • Product overview

    Urea Assay Kit ab83362 is a rapid, simple, sensitive, and reliable assay used to quantify urea in a variety of samples such as serum, plasma, and urine, etc. It can be used as a blood urea nitrogen assay kit (BUN assay kit).


    In the urea assay protocol, urea is acted on by enzymes to form a product that reacts with a probe to generate color (ODmax=570nm). The absorbance is directly proportional to the concentration of urea in the solution. The kit can detect as low as 0.5 nmol per well or 10 µM of urea.


    Urea assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 60 min at 37ºC
    - analyze with microplate reader

  • Platform
    Microplate reader

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    Converting Enzyme (Lyophilised) Blue 1 vial
    Developer (Lyophilised) Orange 1 vial
    Enzyme Mix (lyophilized) Green 1 vial
    OxiRed Probe in DMSO Red 1 x 200µl
    Urea Assay Buffer WM 1 x 25ml
    Urea Standard (100mM) Yellow 1 x 100µl
  • Research areas
  • Relevance
    Urea is a waste product which is produced in the liver, dissolved in blood (in a concentration of 2.5 - 7.5 mM), and secreted by the kidneys. Urea also plays a very important role in protein catabolism, removal of toxic ammonia from the body, and the countercurrent system which allows for reabsorption of water and critical ions in the nephrons.
  • Alternative names
    • Carbamide
    • Carbonyl diamide
    • Carbonyldiamine

Images

  • ab83362 was used as a BUN assay (Blood urea nitrogen assay) to measure urea levels in 80 female Sprague-Dawley rats (age 10-12 months-old).

  • Urea measured in cell culture medium and control medium (MEM), background signal subtracted (duplicates +/- SD).

    The samples were tested neat; 1:4; 1:16 and 1:64. Only the latter two gave reliable results.

  • Urea measured in mouse tissue lysates (mg of extracted protein), background signal subtracted (duplicates +/- SD).

  • Urea measured in cell lysates, background signal subtracted (duplicates; +/- SD).

  • Urea measured in biological fluids, background signal subtracted (duplicates; +/- SD).

  • Example of Urea standard curve using ab83362

Protocols

References

This product has been referenced in:
  • Stulik L  et al. Preventing lung pathology and mortality in rabbit Staphylococcus aureus pneumonia models with cytotoxin-neutralizing monoclonal IgGs penetrating the epithelial lining fluid. Sci Rep 9:5339 (2019). Read more (PubMed: 30926865) »
  • Bharti D  et al. Comparative analysis of human Wharton's jelly mesenchymal stem cells derived from different parts of the same umbilical cord. Cell Tissue Res 372:51-65 (2018). Functional Studies . Read more (PubMed: 29204746) »
See all 17 Publications for this product

Customer reviews and Q&As

1-10 of 24 Abreviews or Q&A

Works good

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Easy to follow instruction and good assay results..

Abcam user community

Verified customer

Submitted Oct 01 2018

Urea Assay Kit

Excellent Excellent 5/5 (Ease of Use)
Abreviews
perfect regarding the researcher and technician

Abcam user community

Verified customer

Submitted Jan 24 2018

Answer

We have not tested urea that has been reacted at both amine groups. However, it will likely not work because the enzyme reaction is very specific for urea.

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Question
Answer

The phenol red could interfere if the amount of medium used is enough to contribute red color to the sample. Typically when diluted medium is used and the volume is made up to 50ul with buffer, the color does not interfere in the assay. Since the probe emits at 570nm and this is the pink-orange-red zone of the spectrum,red color from the medium can interfere if undiluted medium is used.

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Answer





For all samples, including tissue or cell extracts, add 1-25ul sample to each well and volume up to 50ul with Assay Buffer.

Add 50 ul Urea Reaction Mix to each sample or standard.

Final volume in each well will be 100ul.


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Answer


To answer your questions:

1) The desired wavelength for reading the samples is 570 nm.

If the detection band width on the plate reader is ± 30, then a wavelength of ± 30 of 570 can be used, which is 540-600 nm. Otherwise the efficiency of detection of this assay will decrease.
However, from an earlier customer inquiry, the lab replied that they would not recommend reading this assay at 590 nm.

This would then also apply to reading the assay at 450 or 620 nm. This would not be recommended as you might not get a signal at these wavelengths.


2) The kit datasheet described what the kit contains (https://www.abcam.com/ab83362.html)

The protocol lists the additional materials that is required as, but not provided:
Microcentrifuge, Pipettes and pipette tips, Fluorescent or colorimetric microplate reader,  96 well plate, Orbital shaker

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Answer

Thank you for contacting us.

The calculation is right. We suggest in the protocol diluting the standard to get 0.5milli molar per 1000ul.

This 0.5 is a molar solution means, it is 0.5 milli mol per litre per 1000ul i.e. 0.5 nano mol per micro litre.

so 2ul of this solution would be 1nmol/well

4ul would be 2nmol/well and so on.

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

Thank you for contacting us.

I have double checked your question with the laboratory, and I am happy to confirm, that this kit should work with the buffers your client mentions. It will work with all common lab buffers. The only ones it would not work with are the ones which can inactivate the enzymes in this assay kit. Since we cannot disclose all the components in such mixtures, your client will need to clarify it with you/us before changing the buffers.

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

Thank you for your enquiry regarding ab83362.
If the detection band width on the plate reader is ± 30, then a wavelength of ± 30 of 570 can be used, which is 540-600 nm. Otherwise the efficiency of detection of this assay will decrease.
Unfortunately, we not have data showing that hemolysis would affect the reading, but I would think that a cleaner serum sample would give better results.
There is no exact amount of mouse serum which will be right for this assay. For unknown samples, we suggest testing several doses of your sample to ensure the readings are within the standard curve range. I would suggest making a dose curve initially to determine the optimal starting sample volume.
I hope this will be useful for you.

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Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange a refund or credit note in compensation.

I look forward to hearing from you with details of how you would like to proceed.

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1-10 of 24 Abreviews or Q&A

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