Heparin-Sepharose 6FF

Heparin-agarose Gel 6FF Product Description:

Heparin is an acidic polysaccharide containing sulfate, which is coupled to an activated agarose gel, which has high physicochemical stability. Heparin can bind to anticoagulant factor III, coagulation factor, protein synthesis factor, lipoprotein, interferon, nucleic acid binding protein, restriction endonuclide, thrombin and thrombinoid biological macromolecules, so heparin agarose gel can be used for purification of these substances.

Heparin-Agarose Gel 6FF Product Features :

Heparin-Agarose Gel 6FF Scope of Application

Heparin agarose gel is used for the separation and purification of biological macromolecules such as anticoagulation factor III, coagulation factor, protein synthesis factor, lipoprotein, interferon, nucleic acid binding protein, restriction endonuclide, thrombin and thrombinoid.

Heparin-Agarose Gel 6FF Use method

1. Install the column

(1) Bring all reagents and fillers to room temperature. Prepare the initial buffer (balance) and buffer.

(2) Take the amount of gel required according to the size of the column, wash off 20% ethanol, drain it, and prepare a homogenate with the initial buffer (according to the gel: buffer =3:1 ratio).

(3) Wet the inside of the column and the bottom of the column with water or buffer and keep a small section of liquid (the liquid level is slightly higher than the filter membrane), be sure to make the bottom end free of bubbles.

(4) 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 use bubbles. Open the liquid outlet of the column, make the gel settle freely in the column, and connect the top of the column.

(5) 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. Balance the column with a buffer of 2-3 times the column volume.

Step 2 Balance

Let the equilibrium buffer flow through the column at a constant flow rate until the effluent conductance and pH are unchanged.

3. Sample delivery

(1) The sample is prepared with equilibrium liquid, and the turbid sample should be centrifuged and filtered on the column;

(2) Generally use 20-50mM,pH7.4-8.0 buffer, in order to avoid ion exchange interference, the buffer can be appropriately added 50mM-100mM NaCl, the most commonly used is Tris-HCl buffer.

Step 4 Elution

Stage or linear elution can be performed with a buffer of PH 7.4-8.0 plus 0.2-2M NaCl. regeneration

5. Regeneration treatment

Three column bed volumes were washed with 0.1M Tris-HClpH7.0 buffer, then three bed volumes were washed with 0.1M NaOH flow containing 2MNaCl, and finally three bed volumes were washed with 20% ethanol. Washing the filler with acid and alkali for a long time will reduce the adsorption capacity of the filler, so the cleaning should be as short as possible

Heparin-Agarose gel 6FF Note:

1. After the gel is removed from the cold room or refrigerator, it is best to slowly shake it back to room temperature at room temperature, and then install the column to avoid bubbles affecting the column effect.

2, adsorption: 20-50mM,pH7.4-8.0 buffer, in order to avoid ion exchange interference, the buffer can be appropriately added 50mM-100mM NaCl, the most commonly used is Tris-HCl buffer.

3, elution: you can use pH7.4-8.0 buffer solution plus 0.2-2M for stage or linear elution.

4, the sample must be the same as the pH and conductivity of the buffer of the balance column.

5, different samples, adsorption and elution methods are not the same, can be carried out according to relevant literature.

6. Regeneration treatment of heparin agar-gel affinity fillers: First wash 3 column bed volumes with 0.1M Tris-HClpH7.0 buffer, then wash 3 bed volumes with 0.1M NaOH flow containing 2MNaCl, and finally wash 3 bed volumes with 20% ethanol. Washing the filler with acid and alkali for a long time will reduce the adsorption capacity of the filler, so the cleaning should be as short as possible.

7. The preservation condition of the affinity filler is 20% ethanol, 4-8℃.

Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.

Remark：These protocols are for reference only. Solarbio does not independently validate these methods.

Note：

1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.

2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.

3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.