Phospholipase D Assay Kit, BioAssay(TM)

Phospholipase D (PLD) catalyses the hydrolysis of the phosphodiester bond of glycerophospholipids to generate phosphatidic acid and a free headgroup. Abnormalities in PLD expression have been associated with human cancers. This method provides a simple and high-throughput assay for measuring PLD activity. In this assay, PLD hydrolyzes phosphatidylcholine to choline which is determined using choline oxidase and a H2O2 specific dye. The optical density of the pink colored product at 570nm or fluorescence intensity (530/585 nm) is directly proportional to the PLD activity in the sample.\n\nKey Features:\nSensitive. Use 10ul samples. Detection range: Colorimetric Assay: 0.06-10 U/L, Fluorimetric Assay: 0.04-1 U/L.\nSimple and High-throughput: the assay involves addition of a single working reagent and can be readily adapted to high-throughput assays for drug screening.\n\nApplications:\nDirect Assays: phospholipase D in biological samples.\nDrug Discovery/Pharmacology: effects of drugs on phospholipase D metabolism.\n\nKit Contents:\nAssay Buffer: 10ml Dye Reagent: 120ul\nEnzyme Mix: 120ul Substrate: 1.5ml\nCalibrator: 400ul\nStorage conditions. The kit is shipped on ice. Store all components at -20 degrees C. Shelf life of three months after receipt.\nPrecautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information.\n\nColorimetric Assay:\nLiquid samples can be assayed directly. Solid samples should be homogenized in a suitable enzyme buffer prior to assay.\nNote: SH-containing reagents (e.g. b

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SPECIFICATIONS

Catalog Number

P4074-50P

Size

1Kit

References

1. Su W, Chen Q, Frohman MA. (2009). Targeting phospholipase D with small-molecule inhibitors as a potential therapeutic approach for cancer metastasis. Future Oncol. 5(9): 1477-86.\n2. Carman GM, Fischl AS, Dougherty M, Maerker G. (1981). A spectrophotometric method for the assay of phospholipase D activity. Anal Biochem. 110(1): 73-6.\n3. Imamura S, Horiuti Y. (1978). Enzymatic determination of phospholipase D activity with choline oxidase. J Biochem. 83(3): 677-80.