Fluorophore
Site-specific conjugation of IgG with Alexa Fluor® 488, 555 or 647.
GlyCLICK is a site-specific conjugation technology for IgG using Fc glycan remodeling and click-chemistry. The technology generates stable and homogenous antibody conjugates for IgG from several species and subclasses. Fc-glycan remodeling by complete deglycosylation of the antibody allows for site-specific conjugation using robust click-chemistry, resulting in a degree of label (DOL) or antibody-drug ratio (DAR) of 2.0.
GlyCLICK Fluorophore is available for site-specific labeling of 250 μg or 2 mg IgG with Alexa Fluor® 488, 555 or 647, and is suitable for in vitro cell imaging, immunohistochemistry and flow cytometry.
- Site-specific fluorophore-conjugation to IgG with a degree of labeling (DOL) of 2
- Conjugation of IgG with preserved immunoreactivity
- Improved imaging quality in fluorescence microscopy applications
Available Products
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Fluorescence Microscopy
Flow Cytometry
Unit Definition
GlyCLICK Fluorophore contains sufficient material for labeling of 250 μg or 2 mg IgG. For larger amounts or evaluation of the technology, please contact us.
Content and Storage
Components in GlyCLICK Fluorophore:
- GlycINATOR Immobilized
- UDP-GalNAz
- GalT(Y298L)
- sDIBO (Alexa Fluor 488, 555 or 647)
- All buffers needed
- Concentration and desalting columns
GlyCLICK Fluorophore 488: Content should be stored at different temperatures.
GlyCLICK Fluorophore 555: Content should be stored at +4-8°C.
GlyCLICK Fluorophore 647: Content should be stored at +4-8°C.
Instructions
Safety Data Sheet
Certificate of Analysis
FAQ and Support
Popular FAQ
GlyCLICK is available in kit formats with AlexaFluor®488, AlexaFluor®555, AlexaFluor®647 or in the Azide Activation kit format for conjugation of a label of choice. With the fluorescent label being alkyne-modified (carrying for example DIBO, DBCO or BCN) it is possible to combine it with the GlyCLICK azide activation kit.
The final step of GlyCLICK labeling kit is based on a copper free click reaction (SPAAC) and it is a strain promoted reaction between an azide and an alkyne-carrying label. As the name indicates the reaction occurs spontaneously due to the strained bond angles. The use of this click reaction that forms stable conjugates, allows you to freely choose label or payload to combine with your IgG as long as it is alkyne-functionalized. Several alkyne-carrying labels are commercially available with cyclooctyne structures such as DIBO, DBCO or BCN.
All the steps (deglycosylation, azide activation and click reaction) in GlyCLICK are highly efficient. Normally, all molecules are conjugated when using GlyCLICK, resulting in DOL=2. On the application page we have shown by MS-analysis that all molecules have been conjugated and there is no remaining un-conjugated material.
Characterization using LC-MS will give a complete and clear picture of mass shift associated with successful conjugation if the antibody is analyzed reduced or fragmented. LC or LC-MS analysis of HC from a reduced sample or scFc fragments generated by FabRICATOR digestion provides distinct elution peaks for conjugated scFc or mass shifts corresponding to the degree of labelling (DOL) following conjugation. On the application page you can see an example of such analysis. Analysis of fluorescently labeled antibodies can also be performed by absorbance measurements and calculate the DOL using the Extinction coefficient for the antibody and the label and the correction factor for the label.
Support