The experimental design added 20 U, 10 U, 5 U, 2.5 U, 1U and 0.5 U enzymes into the 20 ul reaction system to investigate the effect of Endo S2 on the digestion of human IgG.
M marker
Lan1 human IgG 5μg
Lan2 human IgG 5μg+20 U Endo S2
Lan3 human IgG 5μg+10 U Endo S2
Lan4 human IgG 5μg+5 U Endo S2
Lan5 human IgG 5μg+2.5 U Endo S2
Lan6 human IgG 5μg+1 U Endo S2
Lan7 human IgG 5μg+0.5 U Endo S2
Endo S2
Endo S2
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Product Details
Product Details
Product Specification
| Species | Streptococcus pyogenes |
| Synonyms | Endo-beta-N-acetylglucosaminidase EndoS2、Endoglycosidase S2、EndoS2 |
| Expression System | E.coli |
| Molecular Weight | 93 kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Conjugation | Unconjugated |
| Tag | His Tag |
| Physical Appearance | Liquid |
| Storage Buffer | 20 mM Tris-HCl、50 mM NaCl、pH 7.5 @ 25°C |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | [1] Sjögren Jonathan, et al. "EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans." Glycobiology 10:1053-1063. |
Background
Endo-S2 is an endo-n-acetylglucosaminase derived from streptococcus pyogenes that excises the glycosidic bond between the two innermost n-acetylglucosamines of the glycoprotein n-linking sugar chain. It is effective for high mannose type, hybrid type and double antenna complex type. Can be made for all human IgG, as well as mice, rats, monkey, goat, sheep, cattle and horse IgG antibody glycosyl.
Components
Storage Solution : 200U/ul Endo S2 、20 mM Tris-HCl, 50 mM NaCl. (pH 7.5 @ 25°C) 10*Reaction Buffer： 500 mM sodium acetate (pH 6.0 @ 25°C)
Protocol
1. Combine 100 µg of native IgG, 1 µl of 10*Reaction Buffer and H20 (if necessary) to make a 10 µl total reaction volume.
2. Add 1 µl EndoS2.
3. Incubate reaction at 37°C for 1 hour
Guidelines
Please avoid repeated freeze-thaw cycles.
Unit Definition
One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 5 μg of native mouse monoclonal IgG in 1 hour at 37°C in a total reaction volume of 10 µl.
Picture
Picture
Bioactivity
SDS-PAGE
1μg (R: reducing condition, N: non-reducing condition).
RP-HPLC
