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Cardiolipin Assay Kit (Fluorometric)

The easiest kit in the market to measure Cardiolipin in samples
Catalog #: K944
$625.00

Product Details

Cat # +Size K944-100
Size 100 assays
Detection Method Fluorescence (Ex/Em 340/480 nm)
Species Reactivity Mammalian
Applications This assay provides a convenient and non-ELISA alternative to detect cardiolipin amount in biological samples.
Features & Benefits Simple, fast, does not require lenghty incubation times
Kit Components • CL Assay Buffer
• Probe
• Cardiolipin (5 mM)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Cardiolipin (CL) is a very important phospholipid present in the inner mitochondrial membrane and constitutes about 20% of its total lipid composition. It has a dimeric structure comprised of two phosphatidic acid residues linked by a glycerol bridge. It is essential for several mitochondrial functions such as maintaining activity of the electron transport chain complexes, and other mitochondrial processes including biogenesis, fission, fusion and protein transport. It is involved in apoptosis where it helps alter the mitochondrial membrane structure and aids in the release of cytochrome c. Condition such as diabetes and heart failure are linked to changes in levels of cardiolipin and it is also depleted during aging. Exposure to toxicants like cigarette smoke and organophosphates may cause an alteration in cardiolipin levels and composition, thus adversely affecting health. BioVision’s Cardiolipin Assay 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


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In the sample preparation, how much CL Assay Buffer should be used per cells and how much isolated mitochondria should be used?
For 1 million cells, we recommend using 200 µl of CL assay buffer. You can dilute or add more of the lysate depending on the protein concentration. Homogenize using a dounce homogenizer or mild sonication and please keep samples on ice during homogenization and centrifugation. We tested jurkat cells and the protein concentration used was 9 mg/ml for the cell lysate. We also successfully tested different protein amounts from jurkat cells- 36 ug, 54 ug, 72 ug and 90 ug with K944. For mitochondria, 20- 40 µg of mitochondria should yield a good result.
The figure on the datasheet shows Jurkat cell lysate as an example. May I know the number of cells used and amount of protein you added into the sample well for Jurkat cell lysate to generate this data?
One can use 1 million cells and homogenize in 200 µl of Assay Buffer. Dilute the lysate or add more lysate depending on the protein concentration. Homogenize samples on ice using dounce homogenizer or mild sonication. The protein concentration used was 9 mg/ml and different protein amounts were tested including 36 µg, 54 µg, 72 µg and 90 µg and all of which yielded a good signal.
Does the kit detect the oxidized or the unoxidized form of Cardiolipin?
The assay will not differentiate between the oxidized and unoxidized Cardiolipin. The CL probe binds to both types of Cardiolipin and fluorescence is generated as result of structural changes to the probe on association with Cardiolipin. The probe does not differentiate between the oxidized and unoxidized Cardiolipin.