1. Principle of dialysis bag
Technical indicators: MWCO (molecular weight cut off), unit: Diatoms.
During dialysis, molecules smaller than MWCO permeate the dialysis membrane under the diffusion pressure generated by the difference in concentration of the solution on both sides of the dialysis membrane, and the velocity is proportional to the concentration gradient, membrane area and temperature. For faster dialysis, a smaller diameter dialysis bag can be used to increase the membrane area. Commonly used temperature: 4 ° C, warming up, changing the dialysate outside the bag or using a magnetic stirrer can increase the dialysis speed.
2. Dialysis bag use
The dialysis is to separate the macromolecular proteins with a semi-permeable membrane (dialysis bag). Dialysis is the easiest method to remove salts, small amounts of organic solvents, small biomolecular impurities, and concentrated samples during biomacromolecule preparation.
3. How to use the dialysis bag:
1 Cut the dialysis bag into small pieces of appropriate length (about 10 cm).
2 Dilute 100 times the appropriate amount of dialysis bag treatment solution (Fast-Tre Solution), and place the dialysis bag in it for 10 minutes.
3 Remove the dialysis bag thoroughly with distilled water.
4 If you are not using it, you need to completely soak the dialysis bag in the dialysis bag storage solution (Long-Sav Solution) diluted 100 times and store it at 4 degrees. It is necessary to wear gloves when taking a dialysis bag from this time.
5 Before immersing the dialysis bag in the dialysis bag storage solution, the dialysis bag must be filled with water and then drained, cleaned and rinsed three times.
The experimental requirements are not high, you can use the simple treatment method: you can use it in boiling water for 10 minutes.
6 When using, one end is clamped with the sinking dialysis bag clip. After filling with water, press it with your fingers and check it without leaking before loading the sample. Usually one-third to one-half of the space is left to prevent the water and buffer outside the bag from entering the bag excessively during the dialysis process. After the sample is loaded, the bag is clamped by the floating clip, and a glass bead can be placed in the bag to allow the dialysis bag to be in suspension to perform dialysis. In order to speed up the dialysis, in addition to multiple changes to the dialysate, a magnetic stirrer can also be used. The dialysis container is larger, and large beakers, large tubes, and plastic buckets can be used.
4. Dialysis bag related accessories
5. Precautions and tips in the use of dialysis bags
The dialysis bag must be disposed of before use;
1) Add salt samples to the dialysis bag during salt dialysis, clamp, and dialyze with your choice of preservation solution, usually dialysis overnight at 2-8 degrees, one or two dialysate changes in the middle, and dialysate The volume is as large as possible.
First put the dialysate into the container (the diameter of the container is smaller than the length of the dialysis bag), and then put the clamped dialysis bag vertically, so that the contact area between the dialysis bag and the dialysate is the largest, almost the entire dialysis bag is dialysis When the liquid is in contact, the replacement area of ​​the solution is the largest and the replacement speed is the fastest.
If the clip is stable, then clamp it, put the dialysate into the container (the diameter of the container is smaller than the length of the dialysis bag), and then put the clamped dialysis bag vertically, so that the contact area between the dialysis bag and the dialysate is The largest, almost the entire dialysis bag is in contact with the dialysate, the solution has the largest replacement area and the fastest replacement rate.
If the clip is unstable, try to avoid the end of the clip from contacting the dialysate to avoid leakage. Put the dialysate into the container (the diameter of the container is smaller than the length of the dialysis bag), and then put the clamped dialysis bag flat, so that the contact area of ​​the dialysis bag with the dialysate is not large, and only about half of the dialysis bag is in contact with the dialysate. The replacement speed is the slowest.
If the clip is not particularly unstable, fold the ends of the dialysis bag back and clip it in the middle of the fold back. This will reduce the risk of liquid leakage, and then place the dialysis bag on the liquid level of the container. The clip is up (the diameter of the container is smaller than Dialysis bag length). Be careful to wet the entire dialysis bag at regular intervals, otherwise the dialysis bag that is not exposed to the dialysate may dry out and leak.
The dialysis bag must not be filled because the osmotic pressure of the dialysate is lower than the sample in the dialysis bag. When the salt in the sample goes out, the dialysate will also come in, causing the sample to increase in volume. In severe cases, it may swell the dialysis bag. It can easily leak when the clip is unstable.
Except for the first use, the dialysis bag is still dry. In addition to pretreatment, there is no need to pretreat the second time. Rinse the dialysis bag after it has been used up, then store it at 4 degrees. If the microorganisms are dried or grown, they are discarded, so as not to affect the experimental results.
The time of dialysis is uncertain, but frequent agitation (such as magnetic agitation) and frequent exchange of eluent will speed up the achievement of the goal.
2) The amount of protein adsorption, less than 1 ug / gram, (based on the dry weight of the dialysis bag).
Normal use PH value 5-9 (reusable).
The limit is PH 2-12 (one-time use).
3) Pretreatment: After boiling for 5 minutes, it was washed with distilled water at 60 ° C for 2 minutes, and placed in distilled water at 4 ° C for use.
Long-term preservation solution: 0. 05%-0. 1% sodium azide, 1 mM EDTA or 50% glycerol, temperature (4 ° C), washed with distilled water before use.
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