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Cat. No.: PSAK-200 (for 200T)

Cat. No.: PSAK-1k (for 1000T)

Description

Phosphate Sensor Assay Kit, also known as Phosphate Sensor Assay Kit or Phosphate Fluorescent Assay Kit, is a fluorescent assay kit that uses a fluorescently labeled recombinant phosphate-binding protein to detect inorganic phosphate or the activity of enzymes such as ATPase, GTPase, phosphatase, and phosphodiesterase with ultra-high sensitivity.

This product can detect inorganic phosphate (PO43-) with a sensitivity as low as 100 nM. It is suitable not only for directly detecting inorganic phosphate concentration but also for measuring the activity of enzymes like ATPase, GTPase, phosphatase, and phosphodiesterase that directly or indirectly catalyze the production of inorganic phosphate.

Phosphate Sensor is a chemically labeled product of the E. coli phosphate-binding protein (phosphate-binding protein) A197C mutant recombinant protein with an environmentally sensitive fluorescent group MDCC (N-[2-(1-Maleimidyl)ethyl]-7-(diethylamino)coumarin-3-carboxamide). This inorganic phosphate fluorescent probe is highly sensitive to inorganic phosphate and can quickly and tightly bind to it (Kd~0.1 μM). The binding of inorganic phosphate induces significant conformational changes in the sensor protein, leading to a marked increase in fluorescence intensity. Therefore, this fluorescent probe can be used for rapid, ultra-high sensitivity real-time kinetic detection of inorganic phosphate in solutions.

The fluorescence signal of the Phosphate Sensor significantly enhances upon binding with inorganic phosphate. Under excitation at a wavelength of 430 nm and emission at 450 nm, the fluorescence signal increases approximately 9-fold upon binding with inorganic phosphate.

Components

• Phosphate Sensor (100μM)
• Detection Buffer
• Phosphate Standard (10mM)

Features

• The fluorescence signal of the Phosphate Sensor significantly enhances upon binding with inorganic phosphate. Under excitation at a wavelength of 430 nm and emission at 450 nm, the fluorescence signal increases approximately 9-fold upon binding with inorganic phosphate.
• This product detects inorganic phosphate in the sensitivity range of approximately 100 nM to 5 μM, which is over 100 times more sensitive than the traditional Malachite Green Phosphate Assay.
• The Phosphate Sensor can cause a significant fluorescence enhancement upon binding to inorganic phosphate at concentrations as low as 100 nM, with an observed EC50 of approximately 1.7 μM. Detection data may vary slightly under different conditions.
• This product is easy to operate and user-friendly during the detection process. Typically, 10-30 samples can be measured within 30-60 minutes.

Storage

Store at 20ºC, protected from light, and it remains effective for at least one year.

Precautions

• Due to the high concentration of glycerol in the Phosphate Sensor storage solution, it is recommended to dilute a certain amount of Phosphate Sensor to 10 μM or another suitable concentration with 1X detection buffer according to the amount needed for each experiment to reduce operational errors.
• Please note that the concentration of Phosphate Sensor may vary between batches. Use the concentration indicated in the product manual and on the product label.
• When using a fluorescence microplate reader, to avoid fluorescence signal interference between adjacent wells, it is recommended to use a 384-well black plate with opaque walls between wells.
• Some coated multi-well plates may contain inorganic phosphate, which can interfere with the detection system. Therefore, it is recommended to use uncoated 384-well black plates for detection.
• Due to the varying sensitivity of different fluorescence detection equipment, the final concentration of Phosphate Sensor in the detection system can be adjusted according to the strength of the fluorescence signal during actual detection.
• Avoid introducing bubbles into the detection system. The detection plate can be centrifuged or shaken before detection.
• This product is intended for scientific research by professionals only and is not for clinical diagnosis or treatment, food, or drug use, and should not be stored in ordinary residences.
• For your safety and health, please wear a lab coat and disposable gloves during operation.