製品の概要

  • 製品名

  • 検出方法

    Colorimetric
  • サンプルの種類

    Urine, Serum, Plasma, Other biological fluids, Cell Lysate
  • アッセイタイプ

    Quantitative
  • 全工程の試験時間

    0h 40m
  • 製品の概要

    Glutamate Assay Kit ab83389 provides a sensitive detection method of the glutamate in a variety of samples.


    The glutamate assay kit will only measure free glutamate levels but not glutamic acid found in the backbone of peptides or proteins.


    In the glutamate assay protocol, the enzyme mix recognizes glutamate as a specific substrate leading to proportional color development.The amount of glutamate can therefore be easily quantified by colorimetric spectrophotometry at OD = 450 nm.


    Glutamate assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 30 min at 37ºC
    - analyze with microplate reader

  • 特記事項

    EDTA plasma may not be used with the ab83389 Glutamate Assay Kit. EDTA can interfere with the assay. 

    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

     

  • 試験プラットフォーム

    Microplate reader

製品の特性

  • 保存方法

    Store at -20°C. Please refer to protocols.
  • 内容 ラベル 100 tests
    Glutamate Assay Buffer WM 1 x 25ml
    Glutamate Developer (Lyophilised) Red 1 vial
    Glutamate Enzyme Mix (Lyophilised) Green 1 vial
    Glutamate Standard (0.1M) Yellow 1 x 100µl
  • 研究分野

  • 関連性

    Glutamate, one of the two acidic proteinogenic amino acids, is also a key molecule in cellular metabolism. In humans, glutamate plays an important role both in amino acid degradation and disposal of excess or waste nitrogen. Glutamate is the most abundant swift excitatory neurotransmitter in the mammalian nervous system. It is believed to be involved in learning and memory and has appeared to be involved in diseases like amyotrophic lateral sclerosis, lathyrism, autism, some forms of mental retardation and Alzheimer's disease. Glutamic acid is also present in a wide variety of foods, and has been used as a flavor enhancer in food industry.

画像

  • Glutamate levels of central and peripheral keratocytes were determined using Glutamate assay kit (ab83389). 

  • Example of Glutamate standard curve obtained using ab83389

プロトコール

参考文献

This product has been referenced in:

  • Zhou Q  et al. LincRNA-p21 suppresses glutamine catabolism and bladder cancer cell growth through inhibiting glutaminase expression. Biosci Rep 39:N/A (2019). Read more (PubMed: 30902882) »
  • Zhang K  et al. miR-9 regulates ferroptosis by targeting glutamic-oxaloacetic transaminase GOT1 in melanoma. Mol Carcinog N/A:N/A (2018). Read more (PubMed: 30035324) »
See all 14 Publications for this product

レビューと Q&A

1-10 of 36 Abreviews or Q&A

Glutamate levels in PMN supernatants

Good Excellent 5/5 (Ease of Use)
Abreviews
I wanted to analyze free glutamate in the supernatant of short-term stimulated human blood granulocytes of healthy donors and two groups of clinic patients. The cells were freshly isolated from human whole blood and stimulated for 60-70 minutes at 37 °C in HBSS supplemented with 1.6 mM calcium chloride. Cell-free supernatants were snap-frozen as indicated in the assay protocol and used undiluted in the assay including a HBSS background control. Unfortunately, measured glutamate levels were very low.
The assay protocol is good, the assay is easy and quick to perform.

Abcam user community

Verified customer

投稿 Apr 25 2017

Abreviews
I used lysates of mouse hippocampus. Since the "enzyme" is present in fresh lysates, deproteinization is required
https://www.abcam.com/protocols/deproteinization-protocol
Don't forget to leave aside sample for Bradford normalization before deproteinization

Dr. Tamar Licht

Verified customer

投稿 Mar 08 2016

Question
Answer



I have contacted the lab and they have confirmed that we can add the Glutamate assay buffer to the catalogue to purchase separately to dilute your samples.

However, the lab have also said that you can simply dilute yoursamples in deionized distilled water and then add to the well. Fill up the volume to 50 ul using the assay buffer. This is obviously the cheaper option but if you would like me to add the assay buffer to the catalogue, please let me know and I will do so.

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Answer

Unfortunately neither of these kits will be able to detect glutamic acid in peptides/proteins. ab83389 only measures free glutamate levels not glutamic acid and ab138883 is based on enzymatic reactions so it is most likely that it will not react with the glutamate in peptide or proteins.

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Answer



Non normalement les ions NH3 ne devraient pas interférer le dosage avec ce kit.

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Answer



The Glutamate Assay Kit (ab83389) can be used to asses the level of glutamate in a range of different biological samples. Unfortunately, the dilution range required will need to be assessed for the different samples you are hoping to use the kit with. This is not something we would be able to predict and I do not have any information of these exact samples having been used. You will need to perform a pilot experiment in order to find out at which dilutions the concentration of glutamate falls within the linear range of the standard curve.

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Answer



I've checked with the laboratory, and this kit is very specific for glutamate only. Glutamine is not going to cross react and produce false signals. So, no optimization is required and the glutamine media will be fine.

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Question
Answer

J’ai eu une réponse du laboratoire concernant la préparation de vos échantillons. Les composants de votre tampon ne devraient pas causer d’interférence avec ce kit. Cependant, le laboratoire recommande de diluer vos échantillons dans le tampon qui vient avec le kit puisque il contient des éléments essentiels qui sont nécessaire pour la réaction et qui ne sont pas présent dans votre tampon.

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Answer

Thank you for contacting us.

After doing some research on this subject it seems that astrocytes release glutamate in a Ca2+ dependent manner. It is a very well-known function/property of astrocytes involving transmitter uptake and release.

Here is the wiki page explaining this phenomenon: http://en.wikipedia.org/wiki/Astrocyte

I also add the link to a couple of papers explaining this phenomenon:

http://www.pnas.org/content/97/15/8629

http://www.sciencedirect.com/science/article/pii/S0197018603002900

The kit (ab83389) is very specific in detecting only glutamate since it utilizes a glutamate specific substrate.

You may need to change certain conditions in the cell media and play around with the incubation times; some glutamate uptake as well should also be seen.

I hope this helps. Please do not hesitate to contact us for further assistance.

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Question
Answer

Thank you for contacting us.
Though these kits has not been tested with rat samples however based on the material used in these kits these will compatible with all type of mammalian cells.
I can confirm that these kits will be suitable for Rat samples as well.
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 36 Abreviews or Q&A

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