ConA Agarose
MCE ConA agarose is prepared by covalently coupling ConA to agarose and is widely used for the separation and purification of glycoproteins, membrane proteins, glycolipids, polysaccharides, membrane vesicles containing mannose or glucose residues, IgM, and lipoprotein hormones.
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Storage :
4°C, 2 years. Do not dry or freeze.
- Manual (3996KB)
- COA
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Description & Advantages
Concanavalin A (ConA), also known as Canavalia ensiformis lectin, is a mannose- or glucose-specific binding lectin isolated from the seeds of the jack bean (文字). The molecular weight of the ConA monomer is 26 kDa, and it can bind one Ca2+ and one Mn2+ ion, containing a single sugar-binding site. ConA exists as a homotetramer at alkaline pH (pH > 7.0) and as an activated dimer at acidic pH (pH 4.5 - 5.5). In the presence of Ca2+ and Mn2+, ConA exhibits high affinity for terminal α-D-mannose and α-D-glucose residues.
MCE ConA agarose is prepared by covalently coupling ConA to agarose and is widely used for the separation and purification of glycoproteins, membrane proteins, glycolipids, polysaccharides, membrane vesicles containing mannose or glucose residues, IgM, and lipoprotein hormones.
The specifications of the product correspond to the actual beads volume, with the beads content of 50%.
Protocol
Recommended Buffers (Not Provided)
| Buffer | Composition |
|---|---|
| Binding/Wash Buffer | 20 mM Tris-HCl, 0.5 M NaCl, 1 mM CaCl2, 1 mM MnCl2, pH 7.4 |
| Elution Buffer | Binding/wash buffer containing 0.5 M methyl α-D-mannopyranoside or 0.5 M methyl α-D-glucopyranoside |
| Storage Buffer | 20% ethanol |
Note: Prepare all buffers using ultrapure water and sterilize them by filtration through a 0.45 μm or 0.22 μm membrane. ConA binding activity depends on Ca2+ and Mn2+; avoid EDTA, EGTA, and other metal-ion chelators in samples and buffers.
Sample Preparation
Before loading, centrifuge the sample or filter it through a 0.22 μm or 0.45 μm membrane. If necessary, dilute, dialyze, or desalt the sample with binding/wash buffer so that its pH and ionic strength match those of the equilibration buffer.
Glycoprotein Purification
1. Column Packing: Thoroughly resuspend ConA Agarose, pack it into a suitable chromatography column, and remove the storage solution.
2. Column Equilibration: Equilibrate the column with 5 column volumes of binding/wash buffer. Repeat the procedure 2-3 times.
3. Sample Loading: Apply the sample to the column, allow sufficient contact between the sample and resin, and collect the flow-through. A lower flow rate or repeated loading may improve binding efficiency.
4. Washing: Wash the column with 10-15 column volumes of binding/wash buffer until the UV absorbance baseline stabilizes.
5. Elution: Elute the bound glycoproteins with 5-10 column volumes of elution buffer and collect the eluate in separate fractions.
6. Re-equilibration: Wash the resin with 5-10 column volumes of binding/wash buffer to restore the initial column conditions.
Storage
Wash the column with 5-10 column volumes of ddH2O, equilibrate it with 5 column volumes of storage buffer, and store at 2-8°C.
Storage
4°C, 2 years.
Do not dry or freeze.
Attention
1. Ensure the resin is thoroughly resuspended before use.
2. This product is for R&D use only, not for drug, household, or other uses.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K0272-5 mL | HY-K0272-10 mL | HY-K0272-50 mL |
|---|---|---|---|
| ConA Agarose | 5 mL | 10 mL | 50 mL |
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