Overview

  • Product name

    Coenzyme A Assay Kit (Fluorometric - Green)
    See all Coenzyme A kits
  • Detection method

    Fluorescent
  • Sample type

    Cell culture extracts, Tissue Extracts, Cell Lysate
  • Assay type

    Quantitative
  • Sensitivity

    = 40 nM
  • Assay time

    1h 00m
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Coenzyme A Assay Kit  (Fluorometric - Green) (ab138889) provides an ultrasensitive fluorometric assay to quantitate CoA content by detection of the –SH group in CoA.


    The fluorogenic AcetylCoA green indicator dye used in the kit becomes strongly fluorescent upon reacting with –SH.


    The assay kit can detect as little as 4 pmol CoA/ 100 µL assay volume (40 nM). It can be performed in a convenient 96-well or 384-well microtiter-plate format at Ex/Em =  490/520 nm, and easily adapted to automation without a separation step.


    Coenzyme A assay protocol summary:
    - add samples and standards to wells
    - add reaction mix
    - incubate for 10-60 min whilst measuring fluorescence with a microplate reader

  • Notes

    Previously called Coenzyme A Detection kit  (Fluorometric - Green).

  • Platform

    Microplate reader

Properties

Images

  • CoA dose response was measured in a 96-well black plate with Coenzyme A Detection Kit using a microplate reader. As low as 40 nM (4 pmol/well) of CoA was detected with 30 minutes incubation time (n=3).

Protocols

References

This product has been referenced in:

  • Iuso A  et al. A Homozygous Splice Site Mutation in SLC25A42, Encoding the Mitochondrial Transporter of Coenzyme A, Causes Metabolic Crises and Epileptic Encephalopathy. JIMD Rep 44:1-7 (2019). Read more (PubMed: 29923093) »
  • Iuso A  et al. Mutations in PPCS, Encoding Phosphopantothenoylcysteine Synthetase, Cause Autosomal-Recessive Dilated Cardiomyopathy. Am J Hum Genet 102:1018-1030 (2018). Read more (PubMed: 29754768) »
See all 2 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

The ab138889 CoA detection kit assay buffer is 100 mM Na phosphate buffer, pH = 6. The RIPA buffer may be an issue with the other two kits, since they are enzyme-based, and that kind of assay is generally incompatible with detergents. Your best approach will be to dilute the samples as much as you can with the provided buffers, while maintaining the diluted sample OD values (for the carnitine assay) and RFU values (for the other two) remain within the range of the standards. So, you will need to test serial dilutions of, for instance, 1/2, 1/4, and 1/8.

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Answer

The lab have suggested the follow the below guidelines for sample preparation.

1. For lysis plant cells:
Homogenize the leave with the lysis buffer at 200 mg/mL, and then centrifuge at 2500 rpm for 5-10 minutes, use the supernatant for the assay.

2. For lysis bacterial cells
Collecting bacterial cells by centrifugation ((10,000 x g, 0°C, 15 min). Use about 100 to 10 million cells/mL lysis buffer, leave at room temperature for 15 minutes. And then centrifuge at 2500 rpm for 5 minutes, use the supernatant for the assay.

3. For lysis mammalian cells
Simply remove medium from the plates (wells), use about 100 uL lysis buffer per 1-5 million cells (or 100uL/ well in a 96-well cell culture plate), and leave at room temperature for 15 minutes. You can use the cell lysate directly or simply centrifuge at 1500 rpm for 5 minutes, use the supernatant for the assay.

4. For lysis tissues.
Weight ˜20 mg tissue, wash with cold PBS, homogenize with 400 μl of lysis buffer in a micro-centrifuge tube, and then centrifuge at 2500 rpm for 5-10 minutes, use the supernatant for the assay.

For cells, we suggest starting with ˜50,000-100,000 cells/assay and optimising as is appropriate.

Read More

Answer

Thank you for contacting us. You can simply use freeze-thawed method (20 min each at -80C and RT for 2-3 times). Alternatively, use 0.5% NP40 or Triton to lyse the cells. Please let me know if you have any further questions.

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Answer

Thank you for contacting us.

To our knowledge, ab138889 has not been tested in algae. Theoretically It should work however, we cannot guarantee this. However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting us know you are interested in testing this product.

2. Our scientists will email you an inactive personal discount code for the value of the product.

3. Purchase and test the product at the regular price.

4. Submit your results, including your discount code in the additional notes section of your Abreview.

5. Once the Abreview is submitted, the discount code will become active.

6. Apply your discount code on your next order to receive that value off.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.


If you wish to test this kit on algae cells, we recommend using homogenization, and/or sonication to lyse cells. I am attaching the protocol of the kit to this email so that you can read through all the information about how to use the kit and quantify CoA.

With regards to your other question about a kit for quantifying intra cellular lipid from the algae cell, we do not appear to have such a kit. The only lipid kits available in our catalogue are:

Lipid Peroxidation (MDA) Assay Kit (ab118970)
Hepatic Lipid Accumulation/ Steatosis Assay Kit (ab133131)
Lipid Hydroperoxide (LPO) Assay Kit (ab133085)

I would like to recommend checking the Biocompare website which has an excellent life science products search facility that includes many suppliers. The links are:
www.biocompare.com
http://www.biocompare.com/ProductCategories/2045/Antibodies.html


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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