Ammonia Assay Kit - Modified Berthelot - (Colorimetric) (ab102509)

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Functional Studies - Ammonia Colorimetric Assay Kit (Modified Berthelot Assay) (ab102509)
  • Functional Studies - Ammonia Colorimetric Assay Kit (Modified Berthelot Assay) (ab102509)

Key features and details

  • Assay type: Quantitative
  • Platform: Microplate reader
  • Assay time: 30 min
  • Sample type: Cell culture media, Other biological fluids, Plasma, Serum, Urine
  • Sensitivity: 10 µM

Overview

  • Product name

    Ammonia Assay Kit - Modified Berthelot - (Colorimetric)
    See all Ammonia kits

  • Sample type

    Urine, Serum, Plasma, Other biological fluids, Cell culture media

  • Assay type

    Quantitative

  • Sensitivity

    < 10 µM

  • Assay time

    0h 30m

  • Product overview

    Ammonia Assay Kit - Modified Berthelot (Colorimetric) ab102509 provides a rapid, simple, sensitive, and reliable assay to measure ammonia.


    In this non-enzymatic ammonia assay protocol, ammonia is used to form indophenol, a highly colored product easily quantifiable by colorimetry (λ=670nm) using a plate reader.


    The kit can detect less than 1 nmol(~10 µM) ammonia or ammonium using either a 96 well or 384 well microwell plate.


    Ammonia assay protocol summary:
    - add samples and standards to wells
    - add reagent 1 and reagent 2 and incubate for 30 min
    - analyze with a microplate reader

  • Notes

    Ammonia assay kit ab83360 is an alternative to this product, and is the most popular ammonia assay kit.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components Identifier 100 tests
    Ammonia Reagent 1 Amber 1 x 8ml
    Ammonia Reagent 2 Clear 1 x 4ml
    Ammonium Chloride Standard Yellow 1 x 0.1ml

  • Research areas

  • Relevance

    Ammonia is an important source of nitrogen for living systems. Nitrogen is required for the synthesis of amino acids, which are the building blocks of protein. Ammonia is a metabolic product which is created through amino acid deamination. It plays an important role in both normal and abnormal animal physiology like normal animal acid/base balance.

Images

  • Functional Studies - Ammonia Colorimetric Assay Kit (Modified Berthelot Assay) (ab102509)
    Functional Studies - Ammonia Colorimetric Assay Kit (Modified Berthelot Assay) (ab102509)

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

  • Functional Studies - Ammonia Colorimetric Assay Kit (Modified Berthelot Assay) (ab102509)
    Functional Studies - Ammonia Colorimetric Assay Kit (Modified Berthelot Assay) (ab102509)

    Ammonia measured in biological fluids showing quantity (mM) in tested samples. Samples were diluted 10-1000 fold.

References (2)

Publishing research using ab102509? Please let us know so that we can cite the reference in this datasheet.

ab102509 has been referenced in 2 publications.

  • Raghavan SS  et al. Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli. Microb Cell Fact 18:139 (2019). PubMed: 31426802
  • Wu C  et al. Fluxomic Analysis Reveals Central Carbon Metabolism Adaptation for Diazotroph Azotobacter vinelandii Ammonium Excretion. Sci Rep 9:13209 (2019). PubMed: 31520074

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Question

1. Is pH of the sample an important consideration when using this kit?
2. Can you recommend a protocol for preparing mouse stool samples for use with this kit?

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Answer

Thank you for contacting us.
In response to your questions on this kit, the pH of the sample should be ˜7-8. To use mouse stool sample, add assay buffer to sample. Mix it completely either by pipetting up & down, sonicating or vortexing. Centrifuge to remove insoluble material & use the supernatant to do the assay according to the protocol mentioned in the datasheet.

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

Customer kindly contacted us regarding this product. In particular, they are wondering whether a deproteinizing step would be needed when using this on murine urine samples as protein levels in urine are generally low.

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Answer

Thank you for contacting us. I have spoken with the lab regarding the need to deproteinize mouse urine samples with this kit. They would still strongly recommned this step to attain the best possible data with this product. 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

I was wondering if this kit can be used to measure ammonium (instead of ammonia) and if so, is the procedure the same? What is the detection limit?   Thank you.

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Answer

Thank you for contacting us.

This product may be used to measure ammonium. In thisassay, the standards used are ammonium. Adjustment of the protocol is not required and detection levels will be the same.


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

Product code: 102509
Lot number:
Inquiry: Hello, I am trying to set up the ammonia chloride standard curve using the reagents from your kit (ab102509). I am doing the assay in a standard, clear 96-well plate the reagents at room temperature. My OD readings are a lot lower than those in your example curve- my ODs range from 0.074 (water only background) to 0.213 (10nmol point). Any suggestions? Thanks.

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Answer

Thank you for your inquiry.

The most common reason for a low standard curve is a mistake when preparing the dilutions. I can tell from my on personal experience that a 10 times calculation mistake is especially easy to make.

Therefore I can suggest to check the dilution that were made again.

Also the Ammonia reagents provided with the kit have to be capped tightly when not in use to prevent absorption of ammonia from the air. If this was not done, the efficiency of the reagents decrease and you would see lesser signals.

Please let me know if the problem is solved after re-preparing the standard curve. If the problems persist and the kit was purchased no longer than six month agoI amhappy to offer a free of charge replacement or credit note.

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Question

How long can this product been stored after opening?

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Answer

Thank you for calling Abcam earlier today

I have talked to the lab and the kit components are stable for at least 6 months (and up to 12 month) from the shipping date, if stored at the right temp and conditions.

Please let me know if there is anything else I can help you with.

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Question

We are hoping to detect and quantify the following analytes (all human): Plasma glucose Plasma insulin Serum triglycerides Serum urea Serum creatinine Serum total protein Urinary urea Urinary ammonia At the moment I am trying to pick the most suitable kits for this work.  The factors I am considering are price, assay time, limits of detection, use in published studies. We are interested in purchasing the following Abcam kits: Insulin Human ELISA kit ab100578 Urea Assay kit ab83362 Ammonia Colorimetric kit ab102509 Ammonia Assay kit ab83360 Could you tell me, do you think these kits would be suitable for our work?  And have these kits been used in any published human studies?

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Answer

Thank you for contacting us. The kits you listed are appropriate for your samples, though we recommend ab83360 over ab102509 for assay of ammonia in urine samples. None have been published, to our knowledge. For a quote, we will need a shipping and billing address. Please do not hesitate to contact us if you need any more advice or information.

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Question

Will either of these kits work with serum samples? If so, do you have a sample preparation protocol?

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Answer

Thank you for your inquiry. The lab confirmed that yes, these assay kits are compatible with serum samples from mammalian samples but they did not provide an exact protocol. I've included our general serum sample protocol below: For serum: Collect blood into tube without additive (e.g. BD vacutainer, Cat. No. 8002527) Keep at room temperature for 20 minutes Spin 10 minutes at 3000 rpm Aliquot into small tubes and store at –80C until use. I hope this information helps. Please contact us with any other questions.

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