Phosphatidic Acid Assay Kit (Fluorometric) (ab273335) is a plate-based enzymatic assay for quantitation of Phosphatidic Acid (PA) in cells and tissues.
Fluorescent
Cell culture extracts, Tissue Extracts
Quantitative
Phosphatidic Acid Assay Kit (Fluorometric) (ab273335) is a plate-based enzymatic assay for quantitation of Phosphatidic Acid (PA) in cells and tissues.
Fluorescent
Cell culture extracts, Tissue Extracts
Quantitative
Microplate
Blue Ice
-20°C
-20°C
-20°C
Phosphatidic Acid Assay Kit (Fluorometric) (ab273335) is a plate-based enzymatic assay for quantitation of Phosphatidic Acid (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.
This product is manufactured by BioVision, an Abcam company and was previously called K748 PicoProbeTM Phosphatidic Acid Assay Kit (Fluorometric). K748-100 is the same size as the 100 test size of ab273335.
This supplementary information is collated from multiple sources and compiled automatically.
Phosphatidic acid known also as PA is a lipid molecule involved in cellular signaling. It has a molecular mass of about 478 g/mol. This molecule is found widely within cell membranes serving as a minor but essential phospholipid component. PA performs a mechanical role acting as a precursor for biosynthesis of other lipids. It is also important for protein binding at membranes influencing various signaling pathways within the cell. The expression of phosphatidic acid is variable across different cell types and tissues reflecting its multiple roles in cell function.
PA is important in membrane curvature and vesicle trafficking. It contributes to the balance between membrane bending and tension forces in cells. PA often exists in a complex with proteins such as diacylglycerol kinase and phospholipase D which regulate its levels. These interactions illustrate its dynamic role in cell signaling and structural functions. It plays key roles in mitogenesis and muscle regeneration emphasizing its impact on cellular growth and repair processes.
PA participates significantly in the mTOR signaling pathway and the Ras/Raf/MEK/ERK pathway. Through these pathways it connects with proteins like mTOR and Ras influencing cell metabolism growth and survival. In the mTOR pathway PA regulates nutrient-sensing and energy metabolism. It is vital to cellular responses to growth factors and stress signals coordinating with Ras proteins to manage cell proliferation and apoptosis.
PA is linked to cancer and metabolic syndrome. Its role in mTOR signaling has implications for tumor development and progression making it a target for anticancer therapies. Excessive PA can disrupt metabolic pathways potentially leading to insulin resistance and type 2 diabetes. PA interacts with proteins such as mTOR and reactive oxygen species in these conditions highlighting its impact on disease development. The understanding of PA in these contexts could contribute to targeted therapeutic strategies.
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Example data.
Phosphatidic acid standard curve.
Example data.
Phosphatidic acid content in rat liver and HEK 293 cells. For rat liver extract, 100 mg tissue was homogenized in 1 ml PA assay buffer. Lipid extraction was carried out and was re-suspended in 200 μl 5% Triton X-100 solution. For HEK293, cells were grown to confluency and harvested using a rubber policeman.
Example data.
Assay Specificity: Equimolar amounts (20 nmol) of phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylinositol (PI) and cardiolipin (CL) show interference less than 20% with respect of Phosphatidic Acid. Assay shows specificity for phosphatidic acid.
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