Iptg powder can be used for the local anesthetic action site, it is applied as a spreading agent for post-operative wound pain relief and ulcer pain. Molecular biological reagents commonly used in blue and white spot screening and IPTG induced protein expression in bacteria. IPTG, full name isopropyl- β- D-thiogalactoside, CAS 367-93-1, molecular formula C9H18O5S, molecular weight 384.37 Daltons, belongs to small molecule compounds. White or almost white powder, with low solubility in water but better solubility in organic solvents. There are ionic groups in the molecules, so they have a certain degree of conductivity in water. Stable at room temperature, but easy to decompose under high temperature or strong acid and alkali conditions. There is no obvious odor, but slight organic compound odor may be emitted during use. It is a commonly used inducer used to induce protein expression in bacteria. In medical diagnosis, IPTG can serve as a fluorescent probe or chromogenic agent for detecting specific molecules or tissues in samples. By combining with target molecules, IPTG can generate fluorescent signals or color changes, providing strong support for disease diagnosis.
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Morphological |
Crystalline Powder |
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Color |
White |
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Melting point |
105 ° C |
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Boiling point |
350.9 ° C ( rough estimate ) |
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Density |
1.3329 ( rough estimate ) |
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Storage conditions |
2-8 ° C |
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solubility alcohol |
solublesoluble 40 parts of solvent |
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Acidity coefficient ( pKa ) |
13.00 ± 0.70 ( Predicted ) |
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Soluble in water |
50mg / ml |
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Flash point |
197.8 °C |
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Solubility |
1.6g/l |
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Vapor density |
5.21 (vs air) |
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Refractive index |
1.5060 ( estimate ) |
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Iptg powder, Physical parameters , Mp. : 114 ~ 121 ° C , Description of use , Common molecular biology reagents, commonly used in blue-white screening and IPTG-induced protein expression in bacteria. , Packaging specifications , 1G, 5G, 25G, 100G, 1KG , Storage conditions
2-8 °C refrigerated, protected from light , Hazard description , Danger code : Xi , Risk level : R36 / 37 / 38 , Safety level : S23-24 / 25-36.
IPTG is commonly used in cloning experiments that need to induce β-galactosidase activity. It is often used in combination with X-Gal or Bluo-Gal for blue-white screening of recombinant bacterial colonies, which can be induced by Chemicalbook to express lac operon in E. coli. IPTG binds to lacI repressor protein and changes its conformation to prevent the inhibition of lacZ gene encoded by β-galactosidase.
IPTG, full name isopropyl- β-D-thiogalactoside is a commonly used inducer widely used for inducing and regulating protein expression. Due to its unique structure and function, IPTG is usually prepared using Iptg powder synthesis methods in the laboratory.
Synthetic Route
1. Glycoside synthesis:
First synthesis β-D-thiogalactoside is usually synthesized by the method of glycoside synthesis, by condensing galactose with corresponding bases (such as isopropyl). This step requires the use of protective groups to avoid the generation of by-products in subsequent reactions.
2. Phosphorylation reaction:
On the basis of glycoside synthesis, phosphate groups are introduced through phosphorylation reaction to obtain isopropyl group-β-D-thiogalactoside-5'-phosphate ester. This step requires the use of phosphates and corresponding anhydrides or acids.
3. Deprotection group:
By deprotection group reaction, the protective group in the glycoside is removed to obtain the target product isopropyl- β- D-thiogalactoside. This step requires the use of an acid or alkali solution to facilitate the reaction.
Experimental steps:
Prepare reagents and instruments:
Prepare the necessary sugars, bases, protective groups, phosphates, anhydrides or acids, solvents, and other reagents, as well as necessary experimental instruments such as stirrers, thermometers, spectrophotometers, etc.
Synthesis of glycosides:
Heat and stir galactose with corresponding bases in a solvent, add a catalyst, and promote the condensation reaction. This step requires strict control of temperature and stirring time to ensure the smooth progress of the reaction.
Phosphorylation reaction:
The synthesized glycosides are heated and stirred with phosphate, anhydride, or acid in a solvent, and a catalyst is added to promote the phosphorylation reaction. This step requires controlling the temperature and stirring time, while paying attention to the polarity and dosage of the solvent to ensure the progress of the reaction and the generation of the product.
Deprotection group:
Remove the protective group from the phosphorylation product in an acid or alkali solution to obtain the target product isopropyl- β- D-thiogalactoside. This step requires controlling the pH value and temperature, while paying attention to the polarity and dosage of the solvent to ensure the progress of the reaction and the generation of the product.
Separation and purification:
The target product is separated from the reaction mixture through column chromatography, recrystallization, and other separation and purification methods. This step requires attention to operating conditions such as temperature, solvent dosage, flow rate, etc. to ensure the purity and yield of the product.
Analysis and identification:
Structural identification of the target product is carried out through analytical methods such as nuclear magnetic resonance and mass spectrometry. This step requires the use of corresponding instruments, equipment, and technical means to confirm the structure and purity of the product.
Iptg powder full name isopropyl- β- D-thiogalactoside is a commonly used inducer widely used for inducing and regulating protein expression.
1. Protein expression induction:
IPTG plays an important role in protein expression induction. It is a widely used inducer that can efficiently induce the expression of specific genes in bacteria, thereby obtaining the desired target protein in a short period of time. Its mechanism of action is to bind to the lacI gene of bacteria, inhibit the activity of its transcriptional regulatory protein, thereby opening the lac operon of bacteria and initiating the expression of target genes. When using lactose operons as promoters for protein expression, inducers are required, and IPTG can act as a lactose analogue to induce the expression of galactosidase in Escherichia coli. It cannot be utilized by cells to achieve sustained expression. IPTG binds to lac | products, causing conformational changes away from lacO, thereby activating transcription. This inducible transcriptional regulation has become a commonly used element in the construction of E. coli expression system vectors.
2. Gene expression regulation:
IPTG plays an important role in gene expression regulation. As an important experimental reagent, it plays an important role in gene expression regulation and protein overexpression experiments. By controlling the addition time and concentration of IPTG, regulation of target protein expression can be achieved. This regulatory effect is based on the binding of IPTG to lac | products in lactose operons, altering their conformation, thereby leaving lacO and further activating transcription. This inducible transcriptional regulation mechanism makes IPTG of great significance in research fields such as gene function, protein interactions, and drug screening.
3. Protein crystallization:
In protein crystallization experiments, IPTG is used as an inducer to promote protein crystallization. Its mechanism of action is to change the conformation of proteins by binding to hydrophobic regions in proteins, thereby promoting aggregation and crystallization between protein molecules. This aggregation and crystallization process is reversible, so protein crystallization can be induced by adding IPTG, or protein crystals can be dissolved by removing IPTG.
In protein crystallization experiments, IPTG is usually added to the protein solution as the final concentration of 1mM. This low concentration of IPTG can avoid excessive induction of proteins, thereby achieving better crystallization effects. At the same time, IPTG can also serve as a molecular chaperone to assist in the correct folding and aggregation of proteins, thereby obtaining more uniform protein crystals.
It should be noted that IPTG does not promote protein crystallization in all cases. Some proteins are not sensitive to IPTG, or due to their inherent structure and properties that are not suitable for crystallization, experimental validation and optimization are needed for different proteins to determine the optimal crystallization conditions and methods.
Adverse reactions
Cytotoxicity
Toxicity to prokaryotic cells
Although IPTG is commonly used to induce gene expression in prokaryotic cells, high concentrations of IPTG may have toxic effects on prokaryotic cells. Research has shown that when IPTG concentration is too high, it can interfere with normal metabolic processes within cells. For example, it may affect the energy metabolism of cells, interfere with pathways such as glycolysis and tricarboxylic acid cycle, leading to a decrease in intracellular ATP production and affecting cell growth and proliferation. In addition, high concentrations of IPTG may also damage the integrity of the cell membrane, increase its permeability, lead to leakage of intracellular substances, and allow harmful substances from the extracellular environment to enter the cell, thereby causing cell death. Experiments have found that when Escherichia coli is cultured in a medium containing high concentrations of IPTG, the growth rate of the bacteria significantly slows down and the number of viable bacteria gradually decreases with the extension of cultivation time. This indicates that high concentrations of IPTG have a certain inhibitory and killing effect on Escherichia coli.
Toxicity to eukaryotic cells
In addition to prokaryotic cells, IPTG may also be toxic to eukaryotic cells. There are certain differences in structure and function between eukaryotic cells and prokaryotic cells, but IPTG may still affect the normal physiological functions of eukaryotic cells through various pathways. On the one hand, IPTG may interfere with signal transduction pathways within eukaryotic cells. The signal transduction within cells plays a crucial regulatory role in processes such as growth, differentiation, and apoptosis. IPTG may alter the intensity and direction of signal transduction by interacting with certain signaling molecules within cells, thereby affecting normal physiological activities of cells.
Toxicity to eukaryotic cells
On the other hand, IPTG may induce oxidative stress response in eukaryotic cells. Oxidative stress refers to an imbalance between intracellular oxidation and antioxidant activity, leading to an increase in reactive oxygen species (ROS) production. Excessive ROS can attack biomolecules such as DNA, proteins, and lipids within the cell, causing cellular damage and functional impairment. Research has found that under certain conditions, IPTG treatment of eukaryotic cells significantly increases intracellular ROS levels, accompanied by a decrease in cell viability and an increase in apoptosis rate. This suggests that IPTG may induce oxidative stress response and exert toxicity on eukaryotic cells.
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