Diacylglycerol Assay Kit (ab242293)
Key features and details
- Detection method: Fluorescent
- Sample type: Cell Lysate
製品の概要
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製品名
Diacylglycerol Assay Kit -
検出方法
Fluorescent -
サンプルの種類
Cell Lysate -
製品の概要
ab242293 measures diacylglycerol (DAG) content in samples by a coupled enzymatic reaction system. First, kinase is used to phosphorylate DAG samples, yielding phosphatidic acid. Next, a lipase is used to hydrolyze phosphatidic acid to glycerol-3-phosphate. Finally, the glycerol-3- phosphate product is oxidized by glycerol-3-phosphate oxidase (GPO), producing hydrogen peroxide which reacts with the kit’s Fluorometric Probe (Ex. 530-560 nm/Em. 585-595 nm).
ab242293 is a simple, fluorometric assay that quantitatively measures total DAG in cell lysates using a 96-well microtiter plate format. Each kit provides sufficient reagents to perform up to 100 assays, including blanks, standards and unknown samples. The kit contains a DAG Standard and has a detection sensitivity limit of ~15 µM.
製品の特性
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保存方法
Store at -80°C. Please refer to protocols. -
内容 100 tests 10X Assay Buffer 1 x 1.5ml DAG Standard 1 x 1ml Enzyme Mixture 3 x 1.75ml Fluorometric Probe 1 x 110µl Kinase Mixture 2 x 1ml Lipase Solution 3 x 1.4ml -
関連性
Diacylglycerols (DAG) are key intermediates in the biosynthesis of many cellular lipids and play a fundamental role in biochemical signaling. Structurally, diacylglycerol is a glyceride consisting of 2 fatty acid chains linked by a central glycerol backbone. Diacylglycerol acts as a precursor to many lipids (e.g. triglycerides, phospholipids); however, DAG also functions as a second messenger signalling lipid, produced through hydrolysis of PIP2 by phospholipase C, which initiates intracellular Ca2+ release and PKC activation. Ultimately, the PKC enzymes are involved with many modifications to normal cellular physiology, affecting hundreds of substrates.
画像
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DAG standard curve was performed according to the Assay Protocol. Background has been subtracted.
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HEK293 and COS-7 lipid extracts were prepared according to the extraction procedure above. DAG samples were tested according to the Assay Protocol (phosphatidic acid background was determined +/- Kinase Mixture). Negative control values (without DAG) have been subtracted.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (4)
ab242293 は 4 報の論文で使用されています。
- Bishop CA et al. Detrimental effects of branched-chain amino acids in glucose tolerance can be attributed to valine induced glucotoxicity in skeletal muscle. Nutr Diabetes 12:20 (2022). PubMed: 35418570
- Krzysztynska-Kuleta OI et al. Melanopsin Signaling Pathway in HEK293 Cell Line with Stable Expression of Human Melanopsin: Possible Participation of Phospholipase C beta 4 and Diacylglycerol. Photochem Photobiol 97:1136-1144 (2021). PubMed: 33977551
- Shu Y et al. Hepatocyte-specific PKCß deficiency protects against high-fat diet-induced nonalcoholic hepatic steatosis. Mol Metab 44:101133 (2021). PubMed: 33271332
- Al Batran R et al. Pimozide Alleviates Hyperglycemia in Diet-Induced Obesity by Inhibiting Skeletal Muscle Ketone Oxidation. Cell Metab 31:909-919.e8 (2020). PubMed: 32275862