Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online

Citrate Synthase Activity Assay Kit (ab119692)

Reviews (1)Q&A (8)References (28)
ELISA - Citrate Synthase Activity Assay Kit (ab119692)
  • ELISA - Citrate Synthase Activity Assay Kit (ab119692)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 3 hr 15 min
  • Sample type: Cell culture extracts, Tissue Extracts
  • Sensitivity: 1.56 µg/ml

Overview

  • Product name

    Citrate Synthase Activity Assay Kit
    See all Citrate synthetase kits

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    1 3 5.45%
    Inter-assay
    Sample n Mean SD CV%
    1 3 8.3%

  • Sample type

    Cell culture extracts, Tissue Extracts

  • Assay type

    Enzyme activity

  • Sensitivity

    1.56 µg/ml

  • Range

    1.56 µg/ml - 100 µg/ml

  • Assay time

    3h 15m

  • Species reactivity

    Reacts with: Cow, Human
    Does not react with: Mouse, Rat

  • Product overview

    Citrate Synthase Activity Assay Kit (ab119692) is used to determine mitochondrial citrate synthase (CS) activity in a sample. The enzyme is immunocaptured within the wells of a microplate, and activity is determined by recording color development of TNB, which is generated in the reaction of citrate synthesis. The reaction product, TNB, absorbs at OD = 412 nm.


    This kit immunocaptures only native citrate synthase from the samples. Using the immunocapture method allows the measurement of citrate synthase activity with simple sample preparation and no need for mitochondrial isolation.


    This kit can be used with whole cell extracts or tissue homogenates.


    Citrate synthase assay protocol summary:
    - add samples to wells
    - incubate for 3 hrs to capture enzyme
    - wash with wash buffer
    - add activity solution
    - analyze with microplate reader for 5-30 min at 20s intervals

  • Notes

    Citrate synthase (CS, O75390) is the initial enzyme of the tricarboxylic acid (TCA) cycle. This enzyme is a 51.7 kDa enzyme that catalyzes the reaction of 2 carbon acetyl CoA with 4 carbon oxaloacetate to form the 6 carbon citrate (EC 2.3.3.1). This enzyme is an exclusive marker of the mitochondrial matrix. Citrate synthase is found in nearly all cells capable of oxidative metabolism.

    Storage: All components are shipped cold. 10X Reaction Reagent mixture is shipped lyophilized. Before use, rehydrate the reaction mixture by adding 1mL base buffer. Vortex the tube thoroughly to dissolve the lyophilized powder to prepare 10X Reaction solution. After hydration unused amounts of the 10X Reaction solution should be stored at -80°C for 6 months. Store all other components at 4°C. This kit is stable for 6 months from receipt.

  • Platform

    Microplate reader

Properties

Images

  • ELISA - Citrate Synthase Activity Assay Kit (ab119692)
    ELISA - Citrate Synthase Activity Assay Kit (ab119692)
    Figure 2. Example activity measurements from titrated of Human heart homogenate.
  • ELISA - Citrate Synthase Activity Assay Kit (ab119692)
    ELISA - Citrate Synthase Activity Assay Kit (ab119692)
    Figure 1. Example activity measurements from titrated HepG2 cells.

References (28)

Publishing research using ab119692? Please let us know so that we can cite the reference in this datasheet.

ab119692 has been referenced in 28 publications.

  • Maunder E  et al. Peak fat oxidation is positively associated with vastus lateralis CD36 content, fed-state exercise fat oxidation, and endurance performance in trained males. Eur J Appl Physiol 122:93-102 (2022). PubMed: 34562114
  • Mao L  et al. Discovery of JND003 as a New Selective Estrogen-Related Receptor a Agonist Alleviating Nonalcoholic Fatty Liver Disease and Insulin Resistance. ACS Bio Med Chem Au 2:282-296 (2022). PubMed: 35874496
  • Dong X  et al. Silencing LINC00294 Restores Mitochondrial Function and Inhibits Apoptosis of Glioma Cells under Hypoxia via the miR-21-5p/CASKIN1/cAMP Axis. Oxid Med Cell Longev 2021:8240015 (2021). PubMed: 34777696
  • Maunder E  et al. Temperate performance and metabolic adaptations following endurance training performed under environmental heat stress. Physiol Rep 9:e14849 (2021). PubMed: 33977674
  • Jourdain AA  et al. Loss of LUC7L2 and U1 snRNP subunits shifts energy metabolism from glycolysis to OXPHOS. Mol Cell 81:1905-1919.e12 (2021). PubMed: 33852893
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Testing the Human Citrate Synthase Activity Kit with bovine samples.

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
This kit was purchased to see if it was compatible with mitochondria isolated from bovine PBMCs. The initial data looks good but more tests are needed to see if bovine samples can be stored at -80C for 6 months like stated in the protocol booklet. The image I attached shows the following data: A1 B1 (Blank), C1 D1 (Fresh Sample A), E1 F1 (Fresh Sample B), G1 H1 (1 month old sample C). There was some variation in the methods used to isolate the mitochondria in the fresh and 1 month old sample as we are still working to improve this part of the experiment. It is unknown if the variation in mitochondria isolation methods or the age of the samples used is responsible for the differences observed graphically.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 27 2016

Question

Solubilize 2*10^7 PBMCs in 1mL extraction buffer. BCA for protein concentration, I have 6 samples. pr conc. range from 0.5 to 1.3 mg/mL using 1x Incubation Buffer to adjust concentrations to 100ug/mL according to the protocol. add dilution series of several samples to test the kit. incubate 3h, wash 2 times, add 100uL fresh mixed 1x activity solution (rehyrate and dilute right before adding to the wells.), read at RT at 412nm for 30 min. NO COLOR change in each well! Any suggestions?

Read More
Answer

1. This kit passed QC test so all the components should be prepared properly.
2. Try using a higher protein concentration (e.g. 500ug/ml in lieu of 100ug/ml), since different cell lines have different expression level of CS.
3. Increase the incubation time of the incubation buffer to O/N rather than 3hr.

This kit works only with human samples. I could not find a proper positive control in our catalog, we tested this kit with the following materials which can be used as positive controls. These are HepG2 cells, HDFn cells (other fibroblast cells should also work), human liver homogenate, and human heart homogenate.

I hope this information is helpful to you and yields improved results. Please do not hesitate to contact us with any further questions.

Read More

Question

Thank you very much for the prompt response and the information on complex I activity.

I also use the following kits:

ab109908 (complex II enzyme activity),

ab109909 (complex IV activity),

ab109716 (ATP synthase activity) and

ab119692 (citrate synthase activity).

Could you please as well let me know which factors should I use to convert them to mM/min instead of mOD/min? I use and will use often these kits for a large study.

Thanks in advance for your great help and best regards

Read More
Answer

Thank you for contacting us.

Same process for converting all to molarity per amount of protein loaded into well.
Note these are all per cm and should be converted into path length for microplate which should be about 0.6cm.

Extinction coefficients

ab109908 (complex II enzyme activity),
ε600 = 21 mM-1cm-1

ab109909 (complex IV activity),
ε550 = 21.1 mM-1cm-1

ab109716 (ATP synthase activity)
ε340 = 6.220 mM-1cm-1

ab119692 (citrate synthase activity).
Ε412 = 14.1 mM-1cm-1

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question

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:

1. For citrate synthase kit (ab119692): Again, what are the
ingredients of each buffer used in this assays, i.e., 20X buffer,
extraction buffer, 10X blocking buffer, base buffer, and 10X reaction
reagent?

2. For ATP synthase kit (ab109714): Could you further elaborate on the
caption of the figure which indicate the chemistry behinds this assay?
Again, what are the ingredients in each buffer used:i.e., buffer,
detergent, reagent mix, and lipid mix? What's the purpose of using
lipid mix?

3. For PDH assay kit (ab109902):What are the ingredients of each
buffer used: i.e., 20X buffer, detergent, 20X reagent mix, 5X
stabilizer, 100X coupler, 100X reagent dye?

I would very much appreciate for your prompt feedback on that,
which helps me to understand the chemistry behinds all these assays
for tuning my protocols. Thank you.

Read More
Answer

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 details of the citrate synthase and PDH system which they are able to. I will attach these to this email.

These products are covered by our Abpromise guarantee. We provide scientific support, replacement or refund should this product not perform as indicated on the datasheet. More information on our Abpromise may be found at the following link:

https://www.abcam.com/index.html?pageconfig=abpromise

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Read More

Question

Could you kindly help me know what is the composition of the extration buffer (P/N 8201093) used in the Citrate Synthase Activity Assay Kit (ab119692)? I would like to know if it's compatible with my samples

Read More
Answer

Thank you for contacting us.

The composition of the extraction buffer (P/N 8201093) is Hepes, NaCl, and a detergent, pH 7.4. Thus, it does not contain any of the other components you are concerned about (Tris-HCl, KPO4, or EDTA).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question

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.

Read More
Answer

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 antibodies ab40793 and ab61762
NAD/NADH: ab65348 (we also havea kit forNADH Dehydrogenase: ab124535)
Isocitrate Dehydrogenase: ab102528
Isocitrate Lyase: no kits or antibodies currently available

I hope this helps, please let me know if you need any additional information or assistance.

Read More

Question

Good Morning,
I have a question regarding the ab119692 kit. The protocol provided by the website has a sample preparation procedure. I was wondering whether I can use the mitochondria samples that I extracted following a protocol that Mitosciences have using lauryl maltoside.
Thanks!



Read More
Answer

Thank you for contacting us.

Yes you can this kit uses lauryl maltoside to extract active enzymes.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question

uses human skeletal muscle biopsies from children (i.e. has limited samples)
How much tissue do I need to startwith?
Is this different for each kit?

Read More
Answer

Thank you for contacting us.

I have heard back from the lab with the following information:

All these assays require different sample concentrations described in each booklet protocol - but in most cases is approximately 10ug/microplate well.
The good news is that normal whole muscle homogenate contains a good robust mitochondria signal so less is required, muscle samples are described in most if not all protocols.
Another good news is that the same preparation for any of the assays is suitable for all others. I would estimate that if a 1 mg of homogenate can be prepared (0.2mL at 5mg/mL).
This would provide enough material for each of the assays when diluted to the specific concentration in each assay.
We would recommend - if contemplating doing multiple assays - to include a mitochondrial sample as positive control such as ab110338 isolated mitochondria (note this is cow source)



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

Does this kit also work with mouse?

Read More
Answer

Thank you for your inquiry.


Unfortunately, this kit does not work in mouse. The capture antibody works in human but not in rat or mouse. Other species have not been tested yet.


This is the only Citrate Synthase Activity Assay Kit we carry. You may want to try http://www.biocompare.com to see if there is another commercially available.


I am sorry that this information could not be more helpful. Please contact us with any other questions.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com