Shop All Phosphatase Assay Kits

RediPlate™ 96 EnzChek™ Serine/Threonine Phosphatase Assay Kit (Invitrogen™)

The RediPlate™ 96 EnzChek® Serine/Threonine Phosphatase Assay Kit provides a fast, simple and direct fluorescence-based assay for detecting serine/threonine phosphatases and their corresponding modulators and inhibitors. Unlike other microplate assays, this kit provides the necessary reagents predispensed into a 96-well microplate. Simply reconstitute the fluorogenic substrate in the assay wells with buffer, add the desired sample to the wells, incubate and then quantitate the fluorescence in any standard fluorescence-based microplate reader. Inhibitors are included in each assay well to ensure that the assay is selective for Ser/Thr PPases- other phosphatases, including tyrosine phosphatase do not significantly react with the substrate. Additional advantages of this assay include compatibility with non-ionic detergents and insensitivity to free phosphate.

EnzChek™ Phosphatase Assay Kit (Invitrogen™)

Using the EnzChek Phosphatase Assay Kit continuous monitoring of phosphatase activity, even at neutral or acidic pH, can be achieved. The kit contains Molecular Probes' patented 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP) substrate (D6567). DiFMUP is about 100 times more sensitive than 4-methylumbelliferyl phosphate (MUP) for the detection of prostatic acid phosphatase at pH 5.5. The kit is perfect for the continuous assay of prostatic acid phosphatase, protein phosphatase 1 or almost any other phosphatase that can be assayed with nonprotein-based substrates such as MUP or 4-nitrophenyl phosphate (PNPP). Each kit contains a sufficient amount of substrate for ~1000 assays, using a reaction volume of 100 µL per assay. Fluorescence (excitation/emission maxima ~358/455 nm) can be measured in a fluorescence microplate reader or a standard fluorometer.

RediPlate™ 96 EnzChek™ Tyrosine Phosphatase Assay Kit (Invitrogen™)

The RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit is a ready-to-use, fluorescence-based microplate assay for detecting tyrosine phosphatases (PTPase) and their corresponding modulators and inhibitors. Each RediPlate 96-well microplate has removable lanes, enabling researchers to perform only as many assays as needed for each experiment or to perform high-throughput analysis.

Phosphate Sensor

The Phosphate Sensor is a highly sensitive reagent for the detection of inorganic phosphate, either in real-time kinetic or end-point mode. This product is a purified form of recombinant E. coli phosphate-binding protein labeled with the fluorophore MDCC, which is sensitive to changes in its environment. Binding of inorganic phosphate by Phosphate Sensor is rapid and tight (Kd ~ 0.1 µM), resulting in a large increase in fluorescence. The Phosphate Sensor detects phosphate in the high-nanomolar to low-micromolar range, which is 100-fold more sensitive than malachite green. This reagent provides you with a sensitive assay for detecting enzymes that produce inorganic phosphate either directly or indirectly through coupled reactions, such as phosphatases, ATPases, GTPases, and phosphodiesterases.

Phospha-Light™ SEAP Reporter Gene Assay System (Invitrogen™)

Phospha-Light™ assay system is a chemiluminescent reporter gene assay for the sensitive detection of secreted human placental alkaline phosphatase used as a reporter protein.

• Detection of a secreted reporter permits analysis without destroying valuable cells.
• Wide dynamic range of alkaline phosphatase assay lets the user measure enzyme level accurately from femtogram to nanogram range.
• Assay sensitivity is 100- to 1,000-fold better than with either the isotopic⁄non-isotopic assays for chloramphenicol acetyl transferase (CAT) or the colorimetric⁄fluorescent assays for b-galactosidase, providing greater sensitivity than competing assays.
• Highly sensitive assay with a wide dynamic range permits detection of high and low levels of reporter without the need to perform numerous sample dilutions.
• Non-radioactive reporter gene assay kit eliminates concerns over use of radioisotopes.
• Assay can be completed in about one hour, providing fast assay turnaround.

Monitor Expression Over Time
Phospha-Light™ assay system allows sensitive detection of secreted alkaline phosphatase (SEAP) reporter protein in cell culture media. The SEAP gene product is a truncated form of human placental alkaline phosphatase. A secreted reporter protein eliminates the need to prepare cell extracts. A population of cells can be monitored over time and remains intact for further experimentation.

Proprietary Reagents for Ultra-Sensitive Detection
Phospha-Light™ assay system is one of the easiest and fastest methods for optimizing transfection efficiency. A combination of proprietary reagents is the key to the highly sensitive detection achievable with the Phospha-Light™ assay. The system consists of CSPD® high-performance alkaline phosphatase substrate, Emerald™ luminescence enhancer, and a unique buffer system designed to specifically inhibit non-placental endogenous alkaline phosphatase activity. The persistent glow of the chemiluminescent signal permits the use of luminometers without the need for injectors for measurement.

Applications
Phospha-Light™ reporter gene assay system has been used widely for reporter gene assays to measure gene expression in established cell lines and in transfected primary cells, including as a gene knockdown⁄RNA interference read-out. Phospha-Light™ reporter gene assay has been used for a wide variety of viral functional assays, including viral gene expression assays, viral replication, viral fusogenicity, virus neutralization and viral-mediated cell-cell fusion, and viral infectivity. Use of the SEAP reporter protein is very enabling for in vivo reporter gene assays, by assaying serum samples from transgenic, transfected or viral vector-infected animals. Phospha-Light reporter gene assay system has been used to measure SEAP levels in sera from transgenic or transfected whole animals, including mouse, rat, marmoset, monkey and pig sera, and in chicken egg allantoic fluid. The mouse SEAP protein (mSEAP) has recently been developed for improved SEAP protein stability in transgenic mice, and the Phospha-Light system has been used for sensitive detection of mSEAP. For these and additional references, please see our bibliography in the Literature⁄Resources section.

For Research Use Only. Not for use in diagnostics procedures.

NovaBright™ Phospha-Light™ EXP Assay Kit for SEAP (Secreted Placental Alkaline Phosphatase) Reporter Gene Detection (Invitrogen™)

The NovaBright™ Phospha-Light™ EXP Assay Kit is a second generation chemiluminescent reporter gene assay for the ultrasensitive detection of secreted placental alkaline phosphatase (SEAP) or the non-secreted placental isozyme PLAP. This assay combines ultrasensitive detection with a dynamic range of 5 orders of magnitude. The assay system uses the high performance alkaline phosphatase substrate, CSPD®, and a proprietary next generation chemiluminescence enhancer and a proprietary buffer system designed to specifically inhibit endogenous non-placental alkaline phosphatase activity to deliver performance surpassing that of prior generations of chemiluminescnt assays and far surpassing colorimetric and fluorescent detection methods. This new kit provides a much simpler procedure than other assays. The assay is homogenious and requires only two solutions both of which are supplied ready-to-use. Prolonged glow light emission kinetics provides a stable light signal for read-time flexibility. The assay signal is measured in a simple luminometer without injectors. SEAP is a reporter protein that is secreted into the cell culture medium and detected by testing aliquots of the medium. Since SEAP is released into the medium, cell lysis is unnecessary. However, the assay also allows the detection placental alkaline phosphase (PLAP) isoenzyme following cell lysis. Sensitive detection of alkaline phosphatase reporter enzyme enables a large number of applications in many areas of life science research, including gene expression, viral function assays, vaccine development, development of viral vectors and gene delivery methods for gene therapy, in vivo gene expression monitoring and novel cellular functional assays. This SEAP assay is one of the easiest and fastest methods for optimizing transfection efficiency. Prior generations of NovaBright™ Secreted Placental Alkaline Phosphatase (SEAP) Enzyme Reporter Gene Chemiluminescent Reporter Gene Detection Kit 2.0 have been used widely for reporter gene assays to measure gene expression in established cell lines and in transfected primary cells, including as a gene knockdown/RNA interference read-out. This assay has been used for a wide variety of viral functional assays, including viral gene expression assays, viral replication, viral fusogenicity, virus neutralization and viral-mediated cell-cell fusion, and viral infectivity . Use of the SEAP reporter protein is very enabling for in vivo reporter gene assays, by assaying serum samples from transgenic, transfected or viral vector-infected animals. The NovaBright™ reporter gene assay system has been used to measure SEAP levels in sera from transgenic or transfected whole animals, including mouse, rat, marmoset, monkey and pig sera, and in chicken egg allantoic fluid. The mouse SEAP protein (mSEAP) has recently been developed for improved SEAP protein stability in transgenic mice, and the NovaBright&trade SEAP assay has been used for sensitive detection of mSEAP . In addition to reporter gene (gene expression) applications, the assay system is used to measure SEAP as a functional reporter for receptor-ligand binding assays with a SEAP-ligand chimera, protease-mediated secretion, and for secretion pathway activity. The NovaBright&trade assay system has also been used for the cellular measurement of non-placental alkaline phosphatase as a biomarker.

ELF™ 97 Endogenous Phosphatase Detection Kit (Invitrogen™)

The ELF® 97 Endogenous Phosphatase Kit can be used to detect phosphatase activity with the ELF® 97 phosphatase substrate that upon hydrolysis produces a bright and photostable yellow-green fluorescent precipitate at the site of enzymatic activity. This fluorescent precipitate has several unique spectral characteristics, including an extremely large Stokes shift, 180 nm that makes it easily distinguishable from endogenous fluorescence.