5-Bromo-4-chloro-3-indoxyl myo-inositol-1-phosphate, ammonium salt

myo-Inositol 1-(5-bromo-4-chloro-3-indoxylphosphate) ammonium salt; X-phos-Inositol; X-IP

5-Bromo-4-chloro-3-indolyl myo-inositol-1-phosphate ammonium is a fluorogenic substrate used in the detection of phosphatidylinositol phosphate kinases. It can be used as a chromogenic substrate for detecting phosphatidylinositol phosphate kinase activity in tissues, cells, and biological fluids. 5-Bromo-4-chloro-3-indolyl myo-inositol 1 phosphate ammonium is also used as a ligand to detect phosphatidylinositol phosphate kinase activity in vivo. This product is of very high purity and quality. 5-Bromo 4 chloro 3 indolyl myo inositol 1 phosphate ammonium can be used in Enzyme Assays, Diagnostics and Cultures Media.

5-Bromo-4-chloro-3-indoxyl myo-inositol-1-phosphate, ammonium salt (X-Gal) is a widely used chemical reagent in biomedical research due to its unique properties. X-Gal is widely used as a substrate for beta-galactosidase, an enzyme that catalyzes the hydrolysis of lactose into glucose and galactose. The production of a blue color after the addition of X-Gal to bacterial or mammalian cells that express beta-galactosidase is a clear indication that these cells are able to turn on the expression of the intended gene, making X-Gal a valuable tool for gene expression studies. In this paper, we will explore the definition and background, physical, and chemical properties, synthesis and characterization, analytical methods, biological properties, toxicity and safety in scientific experiments, applications in scientific experiments, the current state of research, potential implications in various fields of research and industry, limitations, and future directions of X-Gal.

Definition and Background:

X-Gal is a chromogenic substrate that is widely used in molecular biology to detect the presence of beta-galactosidase. The chemical structure of X-Gal consists of two parts: a galactose moiety and an indoxyl moiety. The galactose moiety is linked to the indoxyl moiety via a beta-glycosidic bond, and the 5-bromo, 4-chloro substitution pattern of the indoxyl moiety gives X-Gal its unique properties.

Physical and Chemical Properties:

X-Gal is a white to off-white powder or crystalline solid that is relatively stable and does not decompose under normal storage conditions. The molecular weight of X-Gal is 523.56 g/mol, and the chemical formula is C14H15BrClNO6P(NH4). X-Gal is soluble in water, methanol, dimethyl sulfoxide, and dimethylformamide, and it has a melting point of 230-232°C.

Synthesis and Characterization:

The synthesis of X-Gal involves the reaction of indoxyl-beta-D-galactopyranoside with hydrobromic acid and hydrochloric acid in the presence of inositol-1-phosphate to yield 5-bromo-4-chloro-3-indoxyl myo-inositol-1-phosphate (X-P) as an intermediate. The X-P intermediate is then reacted with ammonium hydroxide to form X-Gal. The purity of X-Gal can be determined by thin-layer chromatography, high-performance liquid chromatography, and NMR spectroscopy.

Analytical Methods:

The most common method for detecting beta-galactosidase activity using X-Gal is to add the reagent to a culture of bacterial or mammalian cells that have been genetically engineered to express this enzyme. The development of a blue color in these cells indicates that beta-galactosidase activity is present. The absorbance of X-Gal at 420 nm can be used to determine the concentration of the reagent in solution.

Biological Properties:

X-Gal is not toxic to bacterial or mammalian cells at the concentrations typically used in molecular biology experiments. Moreover, the blue color produced by the reaction between X-Gal and beta-galactosidase is stable and does not fade over time. This makes X-Gal a useful tool for identifying gene expression patterns in living cells.

Toxicity and Safety in Scientific Experiments:

X-Gal is a relatively safe reagent to use in scientific experiments as it is not toxic to cells at the concentrations used. However, standard safety precautions should be followed when handling X-Gal, including the use of gloves and protective eyewear.

Applications in Scientific Experiments:

The most common application of X-Gal in molecular biology is the detection of beta-galactosidase activity as an indicator of gene expression. X-Gal can also be used to generate colorimetric reporter genes, such as the lacZ gene, which can be used to monitor gene expression in real-time.

Current State of Research:

X-Gal is a well-established reagent in molecular biology, and its properties have been extensively studied. Recent research has focused on the development of more sensitive and specific methods for detecting beta-galactosidase activity in cells.

Potential Implications in Various Fields of Research and Industry:

The ability to detect gene expression patterns in living cells using X-Gal has broad implications in a variety of fields, including molecular biology, biotechnology, and medical research. The development of more sensitive and specific methods for detecting beta-galactosidase activity using X-Gal could lead to new diagnostic and therapeutic strategies for a variety of diseases.

Limitations and Future Directions:

The main limitation of X-Gal is its inability to detect gene expression patterns in real-time. Future research should focus on the development of more sensitive and specific methods for detecting beta-galactosidase activity using X-Gal, as well as the use of X-Gal in combination with other reporter genes to monitor gene expression in living cells in real-time. Other future directions for X-Gal research could include the modification of its chemical structure to enhance its properties or the use of X-Gal in drug discovery and development.

Conclusion:

In conclusion, X-Gal is a valuable tool in molecular biology research that has broad implications in a variety of fields. Its unique properties make it a useful reagent for detecting gene expression patterns in living cells, and its safety profile and stability make it an attractive option for scientific experiments. The development of more sensitive and specific methods for detecting beta-galactosidase activity using X-Gal could lead to new diagnostic and therapeutic strategies for a variety of diseases. Future research should focus on the continued development of X-Gal as a tool for studying gene expression in living cells.