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Storage:RT，2 years
Allyl glucan gel series fillers are allyl glucan and N, N' -methylene bisacrylamide crosslinked copolymer with an average particle size of 50μm,
At the same time, the rigidity of the matrix is increased, ensuring fast flow characteristics and high resolution.
Separation range
5×103 ~ 2.5×105 (globulin)
Storage conditions
The product should be sealed and stored in a ventilated, dry and clean place between 4℃and 25℃(the storage solution is 20% ethanol), and cannot be frozen. The used column is stored at 4℃(20% ethanol), shelf life: 2 years.
Usage
Separation filler used for purification and separation of protein and other substances. This product is for scientific research only and shall not be used for other purposes.
Procedure
1. Install the column
(1) Equilibrate all materials and reagents to the temperature of the chromatographic experiment. The buffer is prepared and all buffers are degassed.
(2) Check all components of the chromatographic column, especially whether the filter, sealing ring, screw plug is tight, and whether the glass tube is clean and complete.
(3) Take the appropriate amount of gel according to the demand, and clean 20% ethanol with deionized water.
(4) Wet the bottom of the column with water or buffer and maintain a small level, be sure to make the bottom of the bubble free.
(5) Use a glass rod to guide the homogenate along the inner wall of the column into the column at one time, taking care not to produce bubbles. Open the liquid outlet of the column, make the gel settle freely in the column, and connect the top of the column.
(6) Open the peristaltic pump, so that the buffer flow rate of 1.33 times the flow rate of use, so that the column bed is stable (pay attention to the pressure does not exceed the maximum pressure of the packing).
2. Balance the chromatographic column for at least 5-10 column volumes before loading the sample until the baseline of the recorder becomes stable (the pH and conductance of the effluent are equal to the pH and conductance of the upper column Buffer).
3. Sample delivery
(1) The sample is prepared with balanced liquid, the sample must be centrifuged or filtered after sampling (0.45um filter membrane), if the sample salt concentration is too large, it needs to be treated before sampling.
(2) The recommended loading volume does not exceed 5% of the column volume.
Step 4 Elution
Eluting with buffer, keep the flow rate and buffer composition unchanged during elution.
Step 5 Regenerate
Generally washed with buffer to balance, can be used again. In some applications, substances such as denatured proteins or liposomes do not elute during regeneration.
The following situations must be cleaned and regenerated:
(1) Increased back pressure;
(2) Color changes at the top of the column;
(3) resolution reduction;
(4) There is a gap between the conversion joint and the gel surface.
If an increase in pressure is observed, the individual filter valves and filter membranes in the line are checked for contamination before starting the column cleaning procedure, and the precipitated proteins, non-specifically bound proteins and lipoproteins are removed by washing the column with 0.1M NaOH.
Precautions
(1) Before loading the sample, the sample must be filtered through the membrane and remove the pigment, otherwise the impurities and pigments will be adsorbed on the filler, affecting the filler
Normal operation. All buffers need to be filtered with a 0.45um filter.
(2) In the process of use, avoid using high concentrations of strong acids and bases, and the concentration of acids and bases should be less than 0.1 moles. The alkali will slow the flow.
(3) Different samples, adsorption and elution methods are different, can be carried out according to the relevant literature.
Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.
