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CAS RN: | Product Number: A3331

AOL-LecBeads [for Fucα(1-6)/α(1-4)/α(1-3)/α(1-2)]


Purity:
Synonyms:
  • AOL-Agarose
  • Aspergillus oryzae lectin (= AOL)-Agarose
Documents:
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Product Information
Product code: A3331
Product name: AOL-LecBeads [for Fucα(1-6)/α(1-4)/α(1-3)/α(1-2)]
Preparation: AOL is covalently coupled with agarose gel
Quantity: 0.5 ml gel/1 ml (1:1, v/v slurry) suspended in PBS, pH6.5
Protein concentration: about 15 mg/ml gel
Binding capacity: >5 mg of glycoprotein (human Lactoferrin) /ml gel
Preservative: 0.02% [w/v] Sodium azide
Stabilizer: None

Description
The AOL is an Aspergillus oryzae recombinant of carbohydrate-binding protein (Lectin) derived from Aspergillus oryzae (The AOL gene is derived from the same origin as the host cell). This lectin preferably binds to [Fucα(1-6)/α(1-4)/α(1-3)/α(1-2)] glycan epitopes.
The LecBeads is a lectin-immobilized agarose which is capable of capturing glycoproteins via binding to its specific epitope(s), and it is prepared by a covalent coupling method between lectin and agarose.
The Lectin-catch method using LecBeads (as shown in Application tab) is available for selective separation of glycoconjugates (glycoproteins, glycolipids, and so on). This AOL-LecBeads (0.5 ml gel) shows a binding capacity of about 2.5 mg or more human Lactoferrin [hLF] through its binding to a terminal [Fucα(1-6)/α(1-3)] glycan epitopes on hLF.

Storage
Store at 2-10°C. (Avoid freezing).

Related Products
L0169 Lectin, Fucose specific (= AOL) from Aspergillus oryzae (5 mg/ml, PBS pH6.5)
A2659 AOL-Biotin Conjugate
R0235 rPSL1a-LecBeads [for Siaα(2-6)Gal]
R0236 rLSL-N-LecBeads [for Galβ(1-4)GlcNAc, poly LacNAc]
R0237 rMOA-LecBeads [for Galα(1-3)Gal]
R0238 rSRL-LecBeads [for GlcNAcβ(1-2)Man, Galβ(1-3)GalNAc]
R0239 rGRFT-LecBeads [for Manα(1-2)Man]

Product Number A3331
Physical State (20 deg.C) Liquid
Storage Temperature 0-10°C
Condition to Avoid Heat Sensitive
Specifications
Appearance White to Almost white clear liquid to cloudy liquid
Concentration(Lowry Methoed) min. 13.0 mg/mL gel
Binding capacity to pass test(min. 5 mg/mL gel, Human Lactoferrin)
Properties (reference)
GHS
Related Laws:
Transport Information:
HS Number 3504009000
Application
The lectin-catch procedure

< Preparation >
Buffers
Binding buffer : PBS pH7.5
Elution buffer : PBS pH7.5 containing 200 mM L-fucose [Product code: F0065] and 0.1% triton-X100 [Product code: P1775]

< Method of the lectin-catch > ※Centrifuge method

Equilibration

  • Prepare the LecBeads in a new tube.
  • Add the Binding buffer into the tube to equilibrate the gel.
  • Centrifuge the tube at 800 ×g for 3 min and discard the supernatant. (If necessary, repeat equilibration with the Binding buffer >10-fold volume of the LecBeads appropriately.)

Binding

  • Concentrate or dilute the glycoprotein sample to moderate concentration (e.g., 1 mg/ml) with the Binding buffer [Input fraction], and apply the prepared sample to the tube in which the LecBeads was preliminary equilibrated.
  • Agitate the tube at 4°C (※Generally, a dissociation constants (Kd value) of lectins are tend to be decreased at lower temperature) for a few hours (or overnight).
  • Centrifuge the tube at 800 ×g for 3 min and remove the supernatant [Through fraction].
  • Wash the LecBeads by 5 (to 10)-fold of the slurry volume with the Binding buffer several times.


Elution

  • Elute the glycoprotein from the LecBeads by agitation with an appropriate volume of the Elution buffer (e.g. same volume of the LecBeads).
  • Agitate the reaction tube at 4°C or R.T. for a few hours (or overnight).
  • Centrifuge the tube at 800 ×g for 3 min and remove the supernatant [Elution fraction].
  • Check the resulting fractions ([Input fraction] [Through fraction] [Elution fraction]) by protein assay and SDS-PAGE. (In some cases, the repetitive elution step with a small volume of the Elution buffer might improve the elution effect. If necessary, repeat elution steps.)
  • The competitive oligosaccharide included in the obtained elution fractions can be removed by ultrafiltration or dialysis using a sufficient amount of buffer.

 

References


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