MALACHITE GREEN PHOSPHATE ASSAY KIT (C005B-478029)

General description

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

Application

Malachite Green Phosphate Assay Kit has been used in phosphate quantification and steady-state kinetics.

Malachite Green Phosphate Assay Kit has been used to measure the liberation of free orthophosphate in the following applications:

• Phosphatase Assays: liberation of phosphate from peptide, protein, or small molecule substrate.
• Lipase Assays: liberation of phosphate from phospholipids
• Nucleoside Triphosphatase Assays: liberation of phosphate from nucleoside triphosphates (ATP, GTP, TTP, CTP etc).
• Quantitation of phosphate in phospholipids, proteins and DNAs, etc.
• Drug Discovery: high-throughput screen for phosphatase inhibitors.

Features and Benefits

Compatible with high-throughput handling systems.

Suitability

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

Principle

Malachite Green is a basic dye belonging to the triphenylmethane group. 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 quantification of the green complex formed between Malachite Green, molybdate and free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600 - 660 nm) or on a plate reader. The non-radioactive colorimetric assay kits have been optimized to offer superior sensitivity and prolonged shelf life. The assay is simple and fast, involving a single addition step for phosphate determination. Assays can be executed in tubes, cuvettes or multi-well plates. The assays can be conveniently performed in 96 and 384 well plates for high throughput screening of enzyme inhibitors.