Phosphatidic Acid Assay Kit (Fluorometric) (BN00963)

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SKU:
BN00963
  • Phosphatidic Acid Assay Kit (Fluorometric) (BN00963)
  • Phosphatidic Acid Assay Kit (Fluorometric) (BN00963)
  • Phosphatidic Acid Assay Kit (Fluorometric) (BN00963)
€708

Description

ELISA Kit Technical ManualMSDS

Phosphatidic Acid Assay Kit (Fluorometric)

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. Assay Genie'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

Figure: (a) Phosphatidic acid standard curve. (b) 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 as described above and lipid extract was re-suspended in 200 µl 5% Triton X-100 solution. For HEK293, cells were grown to confluency and harvested using a rubber policeman. Extract was prepared as described above. (c) 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. Experiments were performed using kit protocol.

Key Information Description

Product SKU

BN00963

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 and Benefits

Simple, fast, does not require lenghty incubation times

Kit Components

  • PA Assay Buffer
  • PA Converter
  • PA Developer
  • PA Enzyme Mix (in DMSO)
  • PA Standard (1 mM)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans

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