Product Details
| abID | ab273335 |
|---|---|
| Cat # +Size | K748-100 |
| Size | 100 assays |
| Kit Summary | • Detection method- Fluorescence (Ex/Em 535/587 nm) • Sample type- Cell and/or tissue lipid extracts • Species reactivity- Mammalian • Applications- This assay provides a convenient and non-ELISA alternative to detect phosphatidic acid amount in biological samples. |
| Detection Method | Fluorescence (Ex/Em 535/587 nm) |
| Species Reactivity | Mammalian |
| Applications | This assay provides a convenient and non-ELISA alternative to detect phosphatidic acid amount in biological samples. |
| Features & Benefits | Simple, fast, does not require lenghty incubation times |
| Kit Components | • PA Assay Buffer • PA Converter • PA Developer • PA Enzyme Mix • PicoProbeTM (in DMSO) • PA Standard (1 mM) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Phospholipids are amphiphilic molecules containing phosphate groups, and are both critical components of biological membranes and important signaling messengers in a number of biological pathways. Phosphatidic acid (PA) is the simplest phospholipid, consisting of a glycerol backbone with fatty acids bonded to carbon 1 and 2, and a phosphate group bonded to carbon 3. PA is a precursor for the synthesis of more complex phospholipids such as phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine that act as structural components of the plasma membrane. Phosphatidic acid itself is present in the plasma membrane lending the membrane a high curvature and a net negative charge. In addition, it is an important signaling lipid. It acts through a G-protein coupled receptor and is involved in growth regulation and mitosis. BioVision’s phosphatidic acid assay kit is a plate-based enzymatic assay for quantitation of PA in cells and tissues. The PA Converter hydrolyzes PA to form an intermediate which, in the presence of a developer and enzyme mix, converts a non-fluorescent probe to a fluorescent product (Ex/Em= 535/587 nm) that can be quantified. The Converter is specific to PA and does not hydrolyze other phospholipids (e.g. those with more complex head groups such as phosphatidylcholine or cardiolipin), allowing direct quantitation of PA. This kit can detect as low as 40 pmol PA per well
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My sample is not dissolving in 5% Triton X-100 solution after evaporating the chloroform in vacuum oven overnight? I tried to sonicate and the dry film did not dissolve. Do you have any suggestions to improve this process?
Please add 5% Triton X-100 to the dry film of lipids. Heat at 50°C and sonicate the samples. Most of the lipids should go into solution with heating and sonication.
Will the kit measure LPA or is it specific to PA?
The kit is specific for PA and will not detect LPA as the enzyme involved to convert LPA to PA is not included in the kit.
