Properties 

Use: sufficient for 2,500 colourimetric tests

Detection method: colourimetric

Storage temperature: 2-8°C

General description

Phosphate is one of the most abundant minerals in the human body, containing about 1% of total body weight. Phosphate homeostasis is maintained by various regulators such as parathyroid hormone (PTH), calcitriol, and phosphating. Maintaining phosphate homeostasis is essential for several biological processes, including energy metabolism, cell signalling, regulation of protein synthesis, skeletal development, and bone integrity. An adequate level of phosphate is required for the process of apoptosis of mature chondrocytes in the growth plate. Phosphate deficiency is associated with bone pathology and clinical disease.

Application

The Malachite Green Phosphate Assay Kit has been used in phosphate quantification and steady-state kinetics. The Malachite Green Phosphate Assay Kit has been used to measure the release of free orthophosphate in the following applications:

• Phosphatase Assays: Phosphate release from peptide, protein, or small molecule substrate.
• Lipase Assays: Phosphate Release from Phospholipids
• Nucleoside triphosphatase assays: release of phosphate from nucleoside triphosphates (ATP, GTP, TTP, CTP, etc.).
• Phosphate quantification in phospholipids, proteins and DNA, etc.
• Drug Discovery: high throughput screen for phosphatase inhibitors.

Features and Benefits

Compatible with high-performance handling systems.

Suitability

Suitable for the determination of phosphate in a variety of biological and environmental samples.

Principle

Malachite green is a basic dye belonging to the group of triphenylmethane. It is used in biology for the staining of microscopic samples, such as plant tissue, yeast, and bacteria. The Malachite Green method is a sensitive method for the detection of released phosphate. The Malachite Green Phosphate Assay Kit is based on the quantification of the green complex formed between malachite green, molybdate, and free orthophosphate.

The rapid colour formation of the reaction can be conveniently measured in a spectrophotometer (600-660 nm) or in a plate reader. Non-radioactive colourimetric assay kits have been optimized to offer superior sensitivity and long shelf life. The assay is simple and fast, involving a single addition step for phosphate determination. Assays can be run in tubes, cuvettes, or multi-well plates. Assays can be conveniently performed in 96- and 384-well plates for high-throughput detection of enzyme inhibitors.