Other Related Information 

About Lectins		About Sugar Chains
What are lectins? Hemagglutination activity assay  How to use biotin-labeled lectin	Use as a mitogen Lectin Agarose  Lectin solubility	Functions of glycans  Glycans and disease

Lectin Agarose 

Methods of usage
The basic operation is the same as for general agarose gel. However, it is important to note
that the ligand is a physiologically active protein.

•Precautions for usage
1. pH : Can be used at pH 5 to pH 8 (but return to pH 7 to 8 after use)
2. Temperature: 4°C to room temperature (lower temperatures tend to increase affinity for lectins)
3. Organic solvents: Not applicable
4. Surfactants : Effects on lectins vary depending on type and concentration (after usage, wash with a buffer without surfactants)
5. Other: Stirring with a magnetic stirrer and freezing the gel may damage the agarose gel

• Procedure of usage  

1. Wash (Wash with the buffer solution containing the above solution) Using a Kiriyama funnel or glass filter, wash with 5 to 10 times the amount of gel using pure water.
2. Buffer exchange
Wash with initial buffer (5 x the amount of the gel), and suspend in the initial buffer.

3. Degassing: Degas by suction or other methods. Boiling is strictly prohibited.

4. Column packing
Gently suspend the degassed gel and pour it into the column. Linear flow rate: 10 cm/hr

5. Conditioning: Run it in the initial buffer (more than 5 x the gel volume)Linear flow rate: 2-10cm/hr

6. Sample application: Filter the sample beforehand and remove any precipitates.

7. Washing: Run the initial buffer solution until the baseline stabilizes. ->Non-adsorbed fraction

8. Elution: Elute using an elution solution (a solution containing hapten sugar, acid, etc.). Elution is performed using a stepwise gradient or linear gradient.

9. Washing: Run a buffer solution containing 0.5M NaCl to remove any substances that are non-specifically bound to the gel. If using the sample continuously, start from 2. Buffer exchange or 5. Conditioning.

10. Storage: Suspend in a buffer solution containing a preservative (0.02% NaN3) and store at 4°C.

By connecting to a UV monitor and recorder, it is possible to accurately separate each peak.However, if you use a pump, be careful because the liquid level will not stop at the top disc.

The height of the gel bed may change after the pump has been in use for a while. A flow rate of 10 cm/hr or less is recommended.
The sugar (hapten sugar) used for elution depends on the specificity of the lectin.
Some sugars are difficult to actually use because they are very expensive or difficult to obtain.

Lectin	Sugar for elution	Concentration
SSA	Lactose	0.2M
WGA	N-Acetyl-D-glucosamine acid	0.2M pH5