Колонка/пластина для обессоливания Sephadex G-25 Инструкция по применению
Product Introduction
Dextran G-25 desalination column/plate mainly uses the molecular sieve function in its gel polymer compounds of dextran with a network structure. Due to the different molecular weight sizes of the separated substances, when the separated substances pass through the G-25 gel, the substances with a large molecular weight (such as proteins) are blocked outside the gel particles and flow out through the gap between the gel particles, the process is short and the speed of movement is fast, so the substances with a large molecular weight will flow out of the column/plate first. Substances with a small molecular weight (e.g. salts) can penetrate into the micropores of gel particles, the process is long and slow, and substances with a higher molecular weight take longer to flow out of the gel column/plate, so they are finally washed out, and thus the substances are separated.
The parameters of the Sephadex G-25 series media are shown in the table
Projectт | parametric |
Features | Dirty White Gel |
Average particle size (dry) | 150±100μm 92.8 |
Particle size distribution (45-165μm)% | ≥80 |
Split Range (Portuguese) | 100-5000 |
Separation Range (Globulin) | 1000-5000 |
Water ratio (ml wet glue/g dry powder) | 2.3-2.7 |
Linear Flow Rate(0.3MPA, cm/h) | 697.1 |
Maximum Flow Rate(cm/h) | 150 |
Preservation solution | 0.2M NaAc 20 percent ethanol |
storage conditions | 4'C-30'C |
The model number G indicates the amount of water absorbed per gram of gel, for example, the G-25 gel medium indicates that 2.5 ml of water is absorbed per gram of dry gel, i.e. the gel expansion coefficient, which is also the pore size of the gel particles.
Sephadex G-25 series chromatography media are widely used for buffer exchange, protein desalination, separation of proteins of different molecular weights, etc. The maximum sample volume for desalination and buffering can be up to 30% of the column bed volume.
Workflow
1. Balancing the Speaker/Tablet: Once the speaker/tablet with the preloaded G-25 Sefadex is in an upright position, balance the G-25 desalting column/tablet with a buffer 5 to 10 times the volume of the column layer. (Note: Buffer is the desired buffer to replace, e.g. PBS, deionized water, etc.; be careful, control the flow rate)
2. Sampling: After the gel column/plate is balanced with the buffer, pipet the sample and slowly add it to the sieve on the gel surface, open the bottom cap and allow the sample to slowly enter the gel until it is flush with the gel. (Note: The sample volume at the bottom is usually 10-30% of the column layer volume, the sample viscosity should not be too high, otherwise the service life of the G25 gland will be shortened).
3. Elution: The sample is eluted with an eluting buffer after it has fully entered the gel. (Note: The recommended sample flow rate is about 1-2 ml/min. Elution volume is usually 1.2-2.0 times the sample volume).
4. Purification and regeneration: Some proteins are denatured in saline solution, so it may be necessary to purify these proteins by washing the column bed with 0.2 M NaOH or another non-ionic detergent.
5. Storage: After the experiment, store in 0.02% sodium azide solution or 20% ethanol solution.