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Cardiolipin Assay Kit (ab241036)

Reviews (1) Submit a question References (2)
Cardiolipin standard curve after subtraction of background.
  • Cardiolipin content per mg protein.
  • Specificity of the Cardiolipin Probe.

Key features and details

  • Detection method: Fluorescent

Overview

  • Product name

    Cardiolipin Assay Kit

  • Detection method

    Fluorescent

  • Product overview

    The Cardiolipin Assay Kit (ab241036) is a fluorometric assay that makes use of a proprietary probe that fluoresces on association with cardiolipin but not with any other lipids such as phosphatidylcholine and sphingomyelin, making it highly specific.


    Fluorescence can be recorded at Ex/Em 340/480 nm. The kit includes purified cardiolipin as standard and can detect as low as 0.2 nmol of cardiolipin.  

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K944 Cardiolipin Assay Kit (Fluorometric). K944-100 is the same size as the 100 test size of ab241036.

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Cardiolipin (5 mM) 1 vial
    CL Assay Buffer 1 x 25ml
    CL Probe 1 vial

Images

  • Cardiolipin standard curve after subtraction of background.
    Cardiolipin standard curve after subtraction of background.
    Cardiolipin standard curve after subtraction of background.
  • Cardiolipin content per mg protein.
    Cardiolipin content per mg protein.

    Cardiolipin content per mg protein in Jurkat cell lysate, Jurkat cell mitochondria, Bovine heart mitochondria and Yeast Mitochondria (S. cerevisiae).

  • Specificity of the Cardiolipin Probe.
    Specificity of the Cardiolipin Probe.
    Signal from phosphatidylcholine (PC), phosphatidylserine (PS), sphingomyelin (SM) and cardiolipin (CL) demonstrate the specificity of the Cardiolipin Probe. Known amount of each lipid was added (5 nmol/well). Assay was performed using kit protocol.

References (2)

Publishing research using ab241036? Please let us know so that we can cite the reference in this datasheet.

ab241036 has been referenced in 2 publications.

  • Wang Y  et al. Improvement of obesity-associated disorders by a small-molecule drug targeting mitochondria of adipose tissue macrophages. Nat Commun 12:102 (2021). PubMed: 33397994
  • Zhu Y  et al. System biology analysis reveals the role of voltage-dependent anion channel in mitochondrial dysfunction during non-alcoholic fatty liver disease progression into hepatocellular carcinoma. Cancer Sci 111:4288-4302 (2020). PubMed: 32945042

Customer reviews and Q&As

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Use the Abcam Cardiolipin Assay kit Ab-ab241036 to analyze the Cardiolipin level in the membranes of Bacillus subtilis

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
The Laboratory of Prof. Doron Steinberg 1.11.2021
THE AIM: We wanted to know if we can use the Abcam Cardiolipin Assay kit Ab-ab241036 to analyze the Cardiolipin level in the membranes of various Bacillus subtilis strains.
Below we show the protocol and the results obtained. The results of the kit was
compared to the analysis of the same samples run on TLC.
For the cardiolipin analysis we used 3 strains of Bacillus subtilis (A, B and C).
Cell culture preparation
1. Cells from glycerol stocks were plated onto LB agar plates and incubated overnight at 37°C.
2. Next day one colony of each strain was transferred to 3 ml of LB media and incubated at 37°C with continuous shaking at 200 rpm for 5 hours.
3. Then 1 ml of culture was transferred to 100 ml of fresh LB media in 500 ml flask and in-cubated for 24h at 37°C with continuous shaking at 200 rpm until the late stationary phase.
4. Then the OD600 was measured. For strain A it was 3.44, for strains B and C it was 3.66.
5. Then cells were harvested from 40 ml of the culture by centrifugation, and the supernatant was discarded.
Lipids extraction
1. The bacterial pellets were resuspended in 300 µl of Lysozyme (1 mg/ml) and incubated in a water bath at 37°C for 30 min.
2. Then the samples were subjected to three freeze-thaw cycles: The samples were snap frozen in liquid nitrogen and then transferred back to 37°C water bath 3 times with vigorous vortexing.
3. Proteinase K (100 µg/ml) was added and samples were incubated for 30 min at 37°C.
4. At the end of incubation, the samples were centrifuged for 30 min at 21,000 g at 4°C, and the supernatant discarded.
5. Then the residual pellet (~50 µl) was used for lipid extraction using the Bligh and Dyer method.
6. The residual pellets were resuspended in 300 µl of chloroform : methanol (1:2 v/v) followed by a 5 min sonication on ice.
7. 100 µl of chloroform was added followed by another 5 min sonication on ice.
8. 100 µl of ultrapure water was added and then samples were vortexed and centrifuged for 5 min at 20,000 g.
9. 200 µl of the lower phase with chloroform was transferred to a new tube and evaporated in the hood at 35°C.
10. The dried residues were stored at -80°C freezer and then used for the Abcam Assay CL kit or then solved in 50 µl chloroform and then loaded onto TLC plate.
TLC protocol
1. The whole volume of the samples (~50 µl) were loaded onto the TLC plate (TLC Silica gel 60 F254 20*20 cm) and as a control 10 µg cardiolipin (CL) and 5 µg phosphatidylcholine (PC) were loaded.
2. Then the plate was run in chloroform-methanol–water solution (65:25:4, v/v/v) for 1.5 hour.
3. Then the plate was dried for 5 min and sprayed with the Molybdenum Blue spray reagent (Sigma-Aldrich).
4. The picture of the plate was made after 1 min of spraying.
The data presented in Figures 1 A and C show that the lipid extraction method of Bligh and Dyer worked for our stains, and can be detected both with the TLC method and by using the Abcam CL Assay kit (ab241036). It should be noted that the amount of CL in our samples were higher than the samples used for preparing the calibration curve (Figure 1B). There were no back-ground fluorescence from our samples and the phosphatidylcholine (PC) didn’t provide any sig-nals, causing a fluorescence intensities at the similar levels as background control.
Conclusion: The Abcam CL assay kit can be used for determining the cardiolipin levels in the membranes of Bacillus subtilis.
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.

DR. Ronit Sionov

Verified customer

Submitted Nov 09 2021

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