产品信息

Endo F3 is a highly specific recombinant endoglycosidase which cleaves within the chitobiose core of asparagine-linked fucosylated-biantennary and triantennary complex oligosaccharides from glycoproteins. Endo F3 is tagged with a chitin binding domain (CBD) for easy removal from a reaction and is supplied glycerol free for optimal performance in HPLC and MS intensive methods.

Specificity

产品来源

Cloned from Elizabethkingia miricola (formerly Flavobacterium meningosepticum) and expressed in E. coli.

产品类别:

Endoglycosidases Products,

Proteome Analysis Products

应用:

Expression Systems,

Glycan Sequencing,

Proteomics,

Recombinant Glycoprotein Expression,

Glycoprotein Analysis

• 产品组分信息

  本产品提供以下试剂或组分：

  NEB #	名称	组分货号	储存温度	数量	浓度
  P0771S     -20       Endo F3 P0771SVIAL -20 1 x 0.03 ml 8,000 units/ml   GlycoBuffer 4 B1703SVIAL -20 1 x 1 ml 10 X

• 特性和用法

  单位定义

  One unit is defined as the amount of enzyme required to cleave > 95% of the carbohydrate from 10 µg Porcine Fibrinogen in 1 hour at 37°C in a total reaction volume of 10 µl.

  反应条件

  1X GlycoBuffer 4
  Incubate at 37°C

  GlycoBuffer 4
  50 mM sodium acetate
  (pH 4.5 @ 25°C)

  贮存溶液

  20 mM Tris-HCl
  50 mM NaCl
  5 mM EDTA
  pH 7.5 @ 25°C

  热失活

  65°C for 10 minutes

  分子量

  实际: 38.8 kDa

  单位活性检测条件

  Two fold dilutions of Endo F3 are incubated with 10 µg Porcine Fibrinogen and 1X GlycoBuffer 4 in a 10 µl reaction. The reaction mix is incubated at 37°C for 1 hour. Separation of reaction products are visualized via SDS-PAGE.

• 相关产品

  相关产品

  • p0702-endo-h
  • p0703-endo-hf
  • p0709-pngase-f-glycerol-free-recombinant
  • Rapid 快速 PNGase F
  • p0711-rapid-pngase-f-non-reducing-format
  • Endo S
  • 糖苷内切酶 D

• 注意事项

  1. The optimal pH for the cleavage of fucosylated-biantennary (i.e. porcine fibrinogen) and triantennary glycans (i.e. fetuin) is pH 4.5 (10X GlycoBuffer 4). However, fucosylated-biantennary glycans can also be cleaved at pH 6.0 (10X GlycoBuffer 3).
  2. The enzyme should be used under non-denaturing conditions. Detergents, such as SDS, inhibit Endo F3 activity.
  3. Reactions may be scaled-up linearly to accommodate larger reaction volumes.

操作说明、说明书 & 用法

• 操作说明

  1. Typical Reaction Conditions for Endo F3 Protocol (P0771)

工具 & 资源

• 选择指南

  • Endoglycosidase Selection Chart

FAQs & 问题解决指南

• FAQs

  1. Is Endo F3 tagged?  
  2. What is the difference between PNGase F and Endo F3?
  3. How much Endo F3 should I use to deglycosylate a glycoprotein under native conditions?
  4. What is the preferred substrate for Endo F3?
  5. Do detergents inhibit Endo F3 activity?
  6. What is the optimal pH for Endo F3 activity?
  7. How do I eliminate Endo F3 from a reaction?
  8. What is the binding capacity of the Magnetic Chitin Beads used to remove Endo F3?
  9. Is Endo F3 compatible with downstream analysis such as HPLC and Mass Spectrometry?
  10. What are Glycosidases and their uses?

• 实验技巧

  • Endo F3 is tagged with a chitin binding domain (CBD) for easy removal from a reaction using chitin magnetic beads (NEB #E8036)

  • Endo F3 should be used under non-denaturing conditions. Detergents, such as SDS, inhibit Endo F3 activity.

  • The optimal pH for the cleavage of triantennary glycans (i.e. fetuin) is pH 4.5 (10X Glycobuffer 4).

  • Fucosylated-biantennary glycans (i.e. porcine fibrinogen) can be cleaved at both pH 4.5 (10X Glycobuffer 4) and at pH 6.0 (10X Glycobuffer 3).