Glycerol 3-Phosphate Assays
Glycerol 3-phosphate is an important intermediate in the glycolysis metabolic pathway. Animals, fungi, and plants use glycerol 3-Phosphate to produce ATP. It is used to regenerate NAD+ in brain and skeletal muscle cells. Glycerol 3-phosphate has been linked to lipid imbalance diseases such as obesity.
Measurements of glycerol 3-phosphate using Amplite® Fluorimetric Glycerol 3-Phosphate Assay Kit (Cat# 13837) in 3T3-L1 adipocyte, 3T3-L1 fibroblast and HeLa cell lysates. Cells (1-105) were lysed using ReadiUse™ Mammalian Cell Lysis Buffer (Cat# 20012), and then 50 µL of cell lysate was used as glycerol 3-phosphate containing test samples. Assay was performed following the kit protocol.
Amplite® Glycerol 3-Phosphate Assay Kits provide one of the most sensitive methods for quantifying glycerol 3-phosphate. The kits use Amplite® Red substrate to quantify the concentration of glycerol 3-phosphate, which is related to the production of hydrogen peroxide in the glycerol 3-phosphate oxidase-mediated enzyme coupling reactions. The amount of glycerol 3-phosphate is proportional to the concentration of hydrogen peroxide formed in the enzyme coupling reaction cycle. The kits are an optimized "mix and read" assay that is compatible with HTS liquid handling instruments.
Amplite® Colorimetric Glycerol 3-Phosphate Assay Kit detects as little as 12.5 µM glycerol 3-phosphate in 100 µL assay volume. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. Its signal can be easily read with an absorbance microplate reader at ~576±5 nm.
Amplite® Fluorimetric Glycerol 3-Phosphate Assay Kit detects as little as 41 picomole glycerol 3-phosphate in 100 µL assay volume (0.41 µM). The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. Its signal can be easily read with a fluorescence microplate reader at Ex/Em = ~540/590 nm.