€350.00 – €3,200.00
deGlycIT deglycosylates biantennary Fc-glycans on IgG without residual enzyme in the final preparation
deGlycIT contains spin columns with IgGZERO (EndoS) enzyme immobilized on agarose beads. The immobilized enzyme deglycosylates IgG from several species and subclasses by hydrolyzing biantennary Fc glycoforms after the inner most GlcNAc. deGlycIT is available in spin columns for deglycosylation of 0.5 – 100 mg of IgG without any residual enzyme in the final preparation.
| Application | Antibody Deglycosylation, Afucosylation Assessment, Immunoassays |
|---|---|
| Product Group | |
| Source/Origin | Cloned from S. pyogenes, Produced in E. coli (His-tagged), 112 kDa |
| Product Composition | |
| Digestion Site | |
| Specificity | |
| Substrates | Human IgG1, Human IgG2, Human IgG3, Human IgG4, Cow IgG, Fc-fusion proteins, Goat IgG, Horse IgG, Monkey IgG, Mouse IgG, Rabbit IgG, Rat IgG, Sheep IgG |
| Reaction Conditions | 30 minute reaction at 37°C according to protocol specifications |
| Digestion Buffers | |
| Buffer Compatibility | |
| Inhibitors | |
| Unit Definition | 0.1 ml settled resin deglycosylates ≥ 95% of 0.5 mg Human IgG1 Fc-glycans |
First you could increase incubation time and increase temperature to 37°C.
Another explanation for not complete deglycosylation could be if the antibody has higher amounts of high-mannose, some hybrid glycans or bisected glycans. For these glycans IgGZERO has low or no activity. Especially, high-mannose glycans are particular resistant to digestion.
If longer incubation time and increased temperature does not increase deglycsylation we would recommend you to try our other endoglycosidase GlycINATOR (EndoS2). This enzyme has much higher activity on these types of glycans sometimes seen in m.Abs.
Yes, many buffers are compatible with IgGZERO/DeGlycIT. Please keep pH close to neutral.
