Overview

  • Product name
    Aconitase Assay Kit
    See all Aconitase kits
  • Sample type
    Cell Lysate, Tissue Lysate
  • Assay type
    Enzyme activity
  • Assay time
    0h 40m
  • Product overview

    Abcam's Aconitase Assay Kit is a highly sensitive, simple, direct and HTS-ready colorimetric assay for measuring Aconitase activity in biological samples. In the assay, citrate is converted by aconitase into isocitrate, which is further processed resulting in a product that converts a nearly colorless probe into an intensely colored form with a λmax at 450nm.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Aconitase (aconitate hydratase; EC 4.2.1.3) is an iron-sulfur protein containing an [Fe4S4]2+ cluster that catalyzes the stereospecific isomerization of citrate to isocitrate via cis-aconitate in the tricarboxylic acid cycle, a non-redox-active process. Tissue contains two aconitases, a mitochondrial (m-) and a cytosolic (c-) aconitase. They are related, but distinctly different enzymes and are coded for on different chromosomes. Loss of aconitase activity in cells or other biological samples treated with pro-oxidants has been interpreted as a measure of oxidative damage.

  • Tested applications
    Suitable for: Functional Studiesmore details

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components Identifier 100 tests
    (NH4)2Fe(SO4)2 (Lyophilised) Brown/NM 1 vial
    Assay Buffer WM 1 x 30ml
    Cysteine (Lyophilised) Red 1 vial
    Developer (Lyophilised) Purple 1 vial
    Enzyme Mix Green 1 x 200µl
    Isocitrate Standard (100 mM) Yellow 1 vial
    Substrate (lyophilized) Blue 1 vial
  • Research areas
  • Relevance
    Aconitase (aconitate hydratase; EC 4.2.1.3) is an iron-sulfur protein containing an [Fe4S4]2+ cluster that catalyzes the stereospecific isomerization of citrate to isocitrate via cis-aconitate in the tricarboxylic acid cycle, a non-redox-active process. Tissue contains two aconitases, a mitochondrial (m-) and a cytosolic (c-) aconitase. They are related, but distinctly different enzymes and are coded for on different chromosomes. Loss of aconitase activity in cells or other biological samples treated with prooxidants has been interpreted as a measure of oxidative damage.
  • Cellular localization
    ACO1: Cytoplasmic ACO2: Mitochondrial
  • Alternative names
    • ACO 1
    • ACO 2
    • ACO1
    • ACO2
    • Aconitase 1 soluble
    • Aconitase 2 mitochondrial
    • Aconitate hydratase
    • Aconitate hydratase mitochondrial
    • ACONM
    • ACONS
    • Citrate hydro lyase
    • Cytoplasmic aconitate hydratase
    • EC 4.2.1.3
    • Ferritin repressor protein
    • IRE BP 1
    • IREB1
    • IREBP
    • IREBP1
    • Iron regulatory protein 1
    • Iron responsive element binding protein 1
    • IRP1
    • MGC20605
    • MGC33908
    see all

Applications

Our Abpromise guarantee covers the use of ab83459 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.

Images

  • Functional Studies - Aconitase Assay Kit (ab83459)
    Functional Studies - Aconitase Assay Kit (ab83459)

    Aconitase measured in mouse tissue lysates showing quantity (mU) per mg of tested sample.

    Protein concentration for samples varied from 1.5 mg/mL to 10 mg/mL. Samples were diluted 1-27 fold.

  • Functional Studies - Aconitase Assay Kit (ab83459)
    Functional Studies - Aconitase Assay Kit (ab83459)
    30 minute Aconitase sample test using ab83459.
  • Functional Studies - Aconitase Assay Kit (ab83459)
    Functional Studies - Aconitase Assay Kit (ab83459)
    Isocitrate standard curve using ab83459.

Protocols

References

This product has been referenced in:
  • Nishiki Y  et al. Translational control of inducible nitric oxide synthase by p38 MAPK in islet ß-cells. Mol Endocrinol 27:336-49 (2013). Read more (PubMed: 23250488) »
  • Cummins N  et al. Changes to mitochondrial ultrastructure in optic nerve vulnerable to secondary degeneration in vivo are limited by irradiation at 670 nm. BMC Neurosci 14:98 (2013). Read more (PubMed: 24011177) »

See all 2 Publications for this product

Publishing research using ab83459? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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13 reviews or Q&A

10
Votes: Highest First

Dear Sir or Madam,
I'm using the Aconitase Assay Kit ab83459. Now I want to modify my
experiment and therefore I need to know what concentration
(NH4)2Fe(SO4)2 has in your Kit. I would be really glad, if you can
tell me...
Yours ...

Read More



I can confirm that that concentration of the (NH4)2Fe(SO4)2 is approximately 10 mg/ml.

Inquiry: Hi! What is the deal with adding the 'activation mixture' (cysteine and NH4FeSO4) to activate aconitase? A majority of studies including all references in your data sheet are looking at aconitase inactivation as a marker of ROS. If we activate...

Read More

The reason for needing to activate the aconitase in the sample is as follows:

The Iron sulphur cluster required for Aconitase activity is inactivated in biological samples when the aconitase is exposed to oxidants/superoxides. The way to measu...

Read More

Inquiry: We are using the Abcam Aconitase Assay Kit (ab83459). Could you explain why the assay isn't normalized to the amount of protein being used? Should the data be reflected as aconitase activity/mg protein? Thanks

The mU/ml shows activity of Aconitase per unit volume. Another way of expressing the numbers will be if you perform a protein quantitation assay and instead of the ml volume of sample, uses mg of total protein. In this scenario the final numbers can be...

Read More

Can this kit be used for enzymatic activity of aconitase in Bacteria?

For ab83459, we have optimized this assay protocol for samples of mammalian origin. However, theoretically this assay should work with samples from multiple origins including bacteria. Since we have not done it ourselves, I do not know if the assay pro...

Read More

To whom it may concern,
I have purchased 3 assay kits, which I would like to inquire
further details about the products which however are not thoroughly
indicated in the user manual. I would appreciate for your feedback on
that:
...

Read More

Thank you for contacting us.

I have received word back from the lab regarding your request. Unfortunately the details of the reagents and reactions for these products are proprietary and we cannot share them. They have provided what detail...

Read More

Hello!
Thank you for the reply. I mean the aconitase kit ab83459. There is a formula in the section of data analysis.
Good thing about this kit is that a little amount of tissue is necessary for analysis, in my case it would be brain tissue a...

Read More

Thank you for your interest in our products.

I have been discussing your enquiry with my colleagues in the Lab and would like to share the following information:

Regarding ab109712: Aconitase Enzyme Activity Microplate Assay Kit Read More

I wonder if the ABCAM company sells kits to dosage of the enzymes citrate synthase, aconitase, citrate lyase, NAD + and NADH isocitrate dehydrogenase and isocitrate lyase activitys.

Thank you for contacting us. You may be interested in the following kits:

Citrate Synthase: ab119692
Aconitase: ab109712 or ab83459
CitrateLyase: wedo not currently have a kit available, but weyou may be interested in ourprimary ant...

Read More

How specific the kit is?
What is the discount for purchase of 10 kits?

Thank you for contacting us.
I had chat with the product manager of this assay kit and would like to confirm that we are happy to provide you 100% Abreview discount code for this product. The discount code will be, worth the price you pay for ab83...

Read More

The customer kindly phoned to ask about differences between these two products. They are working with duodenum tissues which are very active in iron absorption and are wondering if this high iron contenct will affect the activity assay as well.

Thank you for contacting us.

I have been in touch with the labs regarding use of ab109712 &ab83459 for duodenumtissue because of its iron aborbence.

In regards to ab109712:Although we have not tested in this tissue type, the...

Read More

Hi, Can your aconitase kit be used on frozen tissue? Thanks! Kind Regards

Thank you for your patient. There are two kits in our catalogue targeting Aconitase: ab83459 and ab109712. Both are compatible with frozen tissues, as long as they are frozen appropriately preserving the enzymatic activity. I hope this in...

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1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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