How to operate glucosamine gel layering.

1. Pre-treatment of the gelthe commercially available goods of the cross-linked glucosaccharide gel are mostly dying particles, which must be fully dissolved before use. The method is to slowly pour the dry gel to be used into 5 to 10 times the deionized water, refer to the relevant data gel swelling time, full immersion, and then use the dumping method to remove the surface suspension of small particles, and decompression pumping to remove the bubbles in the gel suspension, ready to load the column. In many cases, gel swelling can also be carried out using heating boiling method, which not only accelerates the rate of swelling, but also removes contaminated bacteria from the gel while removing bubbles.2. Columnthe choice of the column is generally based on the type of sample isolated and the number of samples. When purifying < a href" > protein , the column bed volume should be 25 to 100 times the sample volume. The removal of salt and free luciferin is about 4 to 10 times the sample volume. The bar is too short to affect the separation effect. The column is longer, the separation effect is good, but the column is too long, the separation time is prolonged, and the sample is diluted too much. The inner diameter of the layering column should also be selected appropriately. The inner diameter is too fine, and a "wall effect" occurs, i.e. the flow rate near the pipe wall is greater than the flow rate of the center affects the separation effect. Therefore, the inner diameter and height of the column should have a certain proportion. It should be 1.5 to 1.25 for desalination and 1.20 to 1.100 for purified proteins.

high-resolution molecular sieve layering, the volume of the sample solution is mainly determined by the volume of internal water (Vi), so high water-absorbing gels such as Sephadex G-200, the total bed volume per mL can be added 0.3 to 0.5 mg solution, the use volume of about 0. O2 times the total volume, while low water-absorbing gels such as Sephadex G-75 have a total bed volume plus solute mass of 0.2 mg per mL and a sample volume of 0. Ol times the total volume.(2) sample method: Like ion exchange column layer analysis, after the gel bed is balanced, sucks away the upper liquid, when the balance liquid is lowered to the surface of the bed, close the flow outlet, add the sample fluid with a dropper, open the flow outlet, so that the sample liquid slowly seeps into the gel bed. When the sample liquid surface is exactly the same as the surface of the gel bed, carefully add a few mL lotion to rinse the pipe wall. Then continue to be dewy with a lot of semen.(3) wasculation: after the addition of samples, the layering bed and the desalination reservoir, detector, division collector and recorder connected, according to the nature of the separated substance, pre-estimated a suitable flow rate, quantitative division to collect fluid outflow, each group of one to several mL. Each member can be qualitatively or quantitatively analyzed using appropriate methods.

for used gels, if stored for a short period of time, as long as repeated washing to remove protein and other impurities, add the appropriate amount" preservatives can be; (Responsible Editor: King Kunlun, Great Han)