€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 | Afucosylation Assessment, Antibody Deglycosylation, 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 |
Yes, IgGZERO/deGlycIT are compatible with most commonly used buffers with pH ranging from 6.0 to 8.0, but the reaction conditions might need to be optimized to ensure optimal deglycosylation.
First you could increase incubation time and increase temperature to 37°C. One explanation for incomplete deglycosylation with deGlycIT could be that the antibody has higher amounts of high mannose, hybrid glycans or bisected glycans. For these glycans IgGZERO has low or no activity. High mannose glycans are particularly resistant to digestion recommend you try our other endoglycosidase GlycINATOR (EndoS2). This enzyme has much higher activity on these types of glycans sometimes seen in mAbs.
Yes, many buffers are compatible with IgGZERO/DeGlycIT. Please keep pH close to neutral.
In both products the enzyme is immobilized on agarose beads for IgG-specific hydrolysis of the Fc glycans to the innermost GlcNAc. The deGlycIT contains immobilized IgGZERO (EndoS) and Immobilized GlycINATOR contains GlycINATOR (EndoS2). The GlycINATOR enzyme has much higher enzymatic activity for high mannose and some bisected and hybrid glycans that can occur on mAbs.
