Sephadex G-15 (CAS 11081-40-6) is a beaded containing a large number of hydroxyl groups, which make it easy to swell in water and electrolyte solutions. The appearance is white microspheres, porous, and has a high specific surface area. This porous structure is conducive to the diffusion and separation of biomolecules in the gel. The particle size range is usually between 40-120 microns, which is suitable for various column chromatography and thin-layer chromatography applications. G-type detran have different crosslinking degrees, so their swelling degrees and fractionation ranges are also different. It is composed of a certain average relative molecular weight of detran and glycerol groups cross-linked with each other in the form of ether bridges, and has a three-dimensional network structure.
It as a kind of gel filter filler with good performance, is widely used in biochemistry, molecular biology, medical research, food industry, environmental protection and other fields.
Medical research applications
1. Immunological research
In the field of immunology research, it can be used for antibody isolation and purification, antigen preparation, and immunohistochemical studies. The filtration chromatography technology can realize the effective separation and purification of antibodies and improve the purity and activity of antibodies. At the same time, it can also be used to prepare antigens, providing important experimental materials for immunological research.
2. Cell biology research
In cell biology research, it can be used for the preparation of reagents for cell culture, cell isolation, and detection of cell apoptosis. Through gel filtration chromatography technology, impurities and pollutants in cell culture reagents can be removed, and the success rate of cell culture and cell activity can be improved. Meanwhile, it can also be used for cell isolation and apoptosis detection, providing important experimental methods for cell biology research.
3. Genetic engineering research
In the field of genetic engineering research, it can be used for the isolation and purification of gene expression products, the preparation of gene recombinant products, and the purification of nucleic acid synthesis reagents. Through filtration chromatography technology, gene expression products can be effectively separated and purified, and the purity and activity of gene expression products can be improved. Meanwhile, it can also be used for the preparation of gene recombinant products and purification of nucleic acid synthesis reagents, providing important experimental materials for genetic engineering research.
Food industry applications
1. Food separation and purification
In the food industry, sephadex G-15 can be used for the separation and purification of food. Through filtration chromatography technology, different ingredients in food can be effectively separated and purified to improve the taste and quality of food. For example, in juice production, it can be used to remove impurities and sediment from the juice, improving its clarity and taste.
2. Preparation of food additives
It can also be used as a raw material for the preparation of food additives. Due to its good stability and thickening properties, it can be used as a thickener, stabilizer, and emulsifier in food, improving the taste and stability of food. For example, in ice cream production, it can be used as a stabilizer to improve the taste and stability of ice cream.
Environmental Protection Applications
1. Wastewater treatment
In the field of environmental protection, it can be used for wastewater treatment. The removal and purification of harmful substances in wastewater can be realized by filtration chromatography technology. For example, in the treatment of heavy metal wastewater, it can be used to remove heavy metal ions from the wastewater, reduce the heavy metal content of the wastewater, and meet environmental discharge standards.
2. Water quality monitoring
It can also be used for water quality monitoring. Through gel filtration chromatography technology, different components in water can be separated and purified, so as to achieve accurate monitoring and evaluation of water quality. This method has the advantages of high sensitivity and good accuracy, and is one of the commonly used methods in water quality monitoring.
Other applications
1. Bioreactor carrier
In bioreactors, it can be used as a carrier material. Due to its good biocompatibility and stability, it can immobilize biocatalysts such as enzymes and cells, improving the efficiency and stability of bioreactors. This method has broad application prospects in the design and optimization of bioreactors.
In the laboratory, it can also be used to prepare various laboratory reagents. For example, in the preparation of ELISA kits, impurities and contaminants in the reagents can be removed, improving the purity and stability of the reagents. At the same time, detran G-15 can also be used to prepare other laboratory reagents, such as cell culture reagents, cell separation reagents, etc.
It also plays an important role in the fields of scientific research and teaching. Through filtration chromatography technology, the separation process and principle of different substances can be intuitively displayed to help students understand and master relevant knowledge. At the same time, it can also serve as an important tool in scientific research experiments, providing strong support for scientific research.
Precautions and operating skills for use
Before use, it needs to be swollen and equilibrated. Swelling is to soak dry powder in distilled water for swelling until the volume of does not change. Balance is achieved by equilibrating the chromatography column with the required buffer solution until the baseline of the recorder becomes stable. These steps are crucial for ensuring its separation efficiency and stability.
2. Sample loading and elution
When sampling, attention should be paid to the concentration and volume of the sample. Generally speaking, the sample size should not be too large to avoid overloading and poor separation efficiency. At the same time, suitable eluent and elution conditions need to be selected based on the properties of the sample and separation requirements during elution. For example, in the desalination process, saline free water can be chosen as the eluent; When separating proteins, it is necessary to choose appropriate buffer solutions and pH values.
After multiple uses, it may be contaminated or its performance may decrease, and regeneration treatment is needed at this time. The regeneration method includes steps such as repeated reverse flushing with water and equilibration with buffer solution. Meanwhile, in order to maintain its performance and stability, it is necessary to store it in a dry, cool, and dark environment, avoiding prolonged exposure to air or adverse conditions such as high temperature and humidity.
The molecular sieve effect principle of detran Sephadex G-15 is a complex but interesting process, which depends on the special structure of and the interaction between molecules. The following is a detailed explanation of the principle of its molecular sieve effect:
G-15 is a kind of gel filter filler with good performance, which is prepared by crosslinking detran and epichlorohydrin. It has a three-dimensional network structure, which provides the basis for its molecular sieve effect. The detran G-15 is insoluble in most solvents and stable in water, salt solution, organic solvent, alkali and weak acid solution. This stability makes it widely used in the separation and purification of biomolecules.
Molecular sieve effect is the basic principle that detran G-15 plays a key role in the separation and purification of biomolecules. When the sample solution containing multiple molecular components slowly flows through the chromatographic column, each molecule is moving in two different ways at the same time in the column: vertical downward movement and non directional diffusion movement.
Due to the large diameter of macromolecules, they are not easy to enter the pores of particles, but can only be distributed between particles.
During the elution process, large molecular substances experience relatively less resistance when moving downwards, resulting in a faster movement speed.
The movement of small molecules:
In addition to diffusing in the gap between particles, small molecules can also enter the micropores of particles, that is, into the phase.
In the process of downward movement, small molecules need to diffuse from one particle to the particle gap and then enter another particle, so as to constantly enter and diffuse.
This complex diffusion process causes the downward movement speed of small molecule substances to lag behind that of large molecule substances.
Due to the difference in diffusion mode and movement speed of macromolecules and small molecules in particles, molecules in the sample can be separated according to molecular weight when flowing through the chromatographic column. Specifically:
The molecules with large molecular weight can only enter into the larger pores in the particle gap due to their large diameter, and their moving distance is relatively short, so they flow out of the chromatographic column first.
Do not use double sided tape to attach light panels to dusty, damp, wallpapered or uneven surfaces such as brick, unfinished wood or rough concrete walls;.
The molecule with the smallest molecular weight can enter almost all the pores in the particles and move the longest distance, so it finally flows out of the chromatographic column.
The phenomenon of separation based on molecular weight is known as the molecular sieve effect. The molecular sieve effect of detran G-15 makes it a powerful tool for the separation and purification of biomolecules.
The molecular sieve effect of detran G-15 has been widely used in the separation and purification of biomolecules, including buffer exchange, desalination, separation of small molecules and removal of small molecules. However, it also has certain limitations. For example, it is difficult to separate molecules with different molecular sizes that fall outside the separation range of gl without changing the type of gel. In addition, the separation effect of gel is also affected by its particle size, cross-linking degree and operating conditions.
Sephadex G-15 Combined with Multidimensional Chromatography: Technological Integration and Application Innovation
Sephadex G-15, as a classic dextran gel filtration medium, plays an important role in the field of biological separation and purification by virtue of its unique molecular sieve effect and chemical stability.
Technology integration with multidimensional chromatography
Combination strategy design
To combine G-15 with multidimensional chromatography, a reasonable separation process should be designed based on the characteristics of the sample:
G-15 is used for desalination and small molecule purification to remove interfering substances such as salt and dye, while concentrating the target product. For example, in peptide purification, G-15 can quickly remove over 90% of the salt and concentrate the sample volume to 1/5 of the original volume.
Secondary separation stage
Inject G-15 eluent into a multidimensional chromatography system (such as 2D-HPLC), and further separate components with similar structures or polarities through a combination of reverse phase chromatography, ion exchange chromatography, or size exclusion chromatography. For example, in metabolomics, samples pre treated with G-15 can be separated into isomers or chiral compounds using 2D-HPLC.
Interface and Switching Technology
The efficiency of multidimensional chromatography is highly dependent on the design of interface devices. Common interfaces include:
Enrich the target component in the G-15 eluent through the capture column, and then inject it into the second chromatography column with reverse flow phase. This interface is suitable for high-sensitivity analysis, but the capture efficiency needs to be optimized to avoid sample loss.
Sample ring interface
Using two sample rings with the same volume to work alternately, achieve online mixing of G-15 eluent and the second mobile phase. The interface structure is simple and easy to operate, but it requires controlling the volume of the sample ring to match the flow rate of the second chromatographic column.
Multiple chromatographic columns are used simultaneously to perform second dimensional separation of G-15 eluent, improving analytical flux. This interface is suitable for large-scale sample screening, but it requires coordinating the separation conditions of each chromatographic column to avoid cross interference.
Method development and optimization
The development of a combination method requires comprehensive consideration of the following factors:
Mobile phase compatibility
G-15 eluent is usually an aqueous buffer, while second dimensional chromatography (such as reverse phase chromatography) requires the use of organic solvents (such as acetonitrile, methanol). Mobile phase transition needs to be achieved through gradient elution or online dilution techniques.
Column efficiency equilibrium
The separation range of G-15 is relatively narrow, and a separation mechanism complementary to the second chromatographic column needs to be selected. For example, the combination of G-15 and reverse phase chromatography can achieve dual dimensional separation of molecular weight and polarity.
Data analysis
The two-dimensional data generated by multidimensional chromatography needs to be analyzed using chemometric methods such as principal component analysis and parallel factor analysis to extract the pure color spectrum and spectral information of the target component.
Frequently Asked Questions
Q: Why does Sephadex G‑15 sometimes show abnormally high baseline absorbance at 280 nm during gel filtration?
+
-
A: Sephadex G‑15 contains small amounts of residual dextran oligomers and cross‑linker fragments. These low‑molecular‑weight leachables can elute near the total volume (Vₜ) region and absorb UV light, leading to elevated baseline or false peaks, especially in sensitive protein or peptide assays.
Q: Can Sephadex G‑15 be used for desalting under high organic solvent conditions?
+
-
A: Limited use. G‑15 swells significantly in water but shrinks or compacts in high concentrations of acetonitrile, methanol, or acetone. This reduces bed volume, increases backpressure, and destroys separation efficiency. It is not suitable for routine desalting in >40% organic mobile phases.
Q: Why is Sephadex G‑15 less suitable for separating small peptides below 100 Da?
+
-
A: Its fractionation range is approximately 150–1,500 Da. Molecules below 150 Da are almost completely included in the gel matrix and co‑elute near Vₜ. Thus, it cannot resolve very small polar molecules such as salts, amino acids, or small peptides below its exclusion limit.
Hot Tags: sephadex g-15 cas 11081-40-6, suppliers, manufacturers, factory, wholesale, buy, price, bulk, for sale, CAS 2192307 05 2, mercury indicator powder, 2 biphenyl 4 yl 4 3 9 9 dimethyl 9H fluoren 2 yl biphenyl 3 yl 6 phenyl 1 3 5 triazine, Pamoic acid, CAS 2408302 78 1, 10 bromobenzo d naphtho 1 2 b thiophene