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CAS RN: | 产品编码: R0238

rSRL-LecBeads [for GlcNAcβ(1-2)Man, Galβ(1-3)GalNAc]

× rSRL-LecBeads [for GlcNAcβ(1-2)Man, Galβ(1-3)GalNAc]

纯度/分析方法:
别名：
  • rSRL-LecBeads [for GlcNAcβ(1-2)Man, Galβ(1-3)GalNAc]
  • rSRL-琼脂糖
  • 重组齐整小核菌凝集素(= rSRL)-琼脂糖
  • rSRL-Agarose
  • Recombinant Sclerotium rolfsii lectin (= rSRL)-Agarose
产品文档：
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基本信息 | 技术规格和文件 | 安全和法规 | 应用实例・文献

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1VIAL
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Product information
Product code : R0238
Product name : rSRL-LecBeads [for GlcNAcβ(1-2)Man, Galβ(1-3)GalNAc]
Preparation : Recombinant SRL is covalently coupled with agarose gel
Quantity : 1 ml gel/2 ml (1:1, v/v slurry) suspended in PBS, pH7.5
Protein concentration : about 15 mg/ml gel
Binding capacity : >7.5 mg of glycoprotein (agalacto human Transferrin) /ml gel
Preservative : 0.02% [w/v] Sodium azide
Stabilizer : None

Description
The rSRL is an Escherichia coli recombinant of carbohydrate-binding protein (Lectin) derived from Sclerotium rolfsii. This lectin preferably binds to [GlcNAcβ(1-2)Man, Galβ(1-3)GalNAc] glycan epitopes by its two distinct carbohydrate-binding sites.
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 rSRL-LecBeads (1 ml gel) shows a binding capacity of about 7.5 mg or more agalacto human Transferrin [agalacto hTF] through its binding to a terminal [GlcNAcβ(1-2)Man] glycan epitope on agalacto hTF.

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

Related Products
R0228 Recombinant Sclerotium rolfsii lectin (= rSRL) expressed in Escherichia coli
R0233 rSRL-Biotin [for GlcNAcβ(1-2)Man, Galβ(1-3)GalNAc]
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]
R0239 rGRFT-LecBeads [for Manα(1-2)Man]
A3331 AOL-LecBeads [for Fucα(1-6)/α(1-4)/α(1-3)/α(1-2)]

产品编码 R0238
外观与形状（20°C） 液体
储存温度 冷藏 (0-10°C)
应避免的情况 加热
技术规格
Appearance White to Almost white clear liquid to cloudy liquid
Concentration(Lowry method) min. 9.0 mg/mL gel
Binding capacity to pass test(min. 5 mg/mL gel, Agalacto-human transferrin)
物性（参考值）
GHS
相关法规
新化学物质备案回执号 B1A232224108
运输信息
监管条件代码(*)
应用
The lectin-catch procedure

< Preparation >
Buffers
Binding buffer : PBS pH7.5
Elution buffer : PBS pH7.5 containing 200 mM D-GlcNAc [Product code: A0092] and 0.1% triton-X100　[Product code: P1775]
(Herein, an example of capturing agalacto-type glycoprotein is shown.)

< 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


文章/手册

TCIMail
[TCIMAIL No.192] Glycoprotein Capturing Reagents: Lectin-Immobilized Agarose "LecBeads"
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