GlyCLICK®
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GlyCLICK® Fluorophore
Site-specific conjugation of IgG with Alexa Fluor® 488, 555 or 647
GlyCLICK is a three-step conjugation technology for site-specific and quantitative conjugation of IgG from several species and subclasses.
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.
Why GlyCLICK® Fluorophore?
- 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
GlyCLICK® Fluorophore 488 250 ug
Site-specific conjugation of 250 𝜇g IgG with Alexa Fluor® 488
GlyCLICK® Fluorophore 488 2 mg
Site-specific conjugation of 2 mg IgG with Alexa Fluor® 488
GlyCLICK® Fluorophore 555 250 ug
Site-specific conjugation of 250 𝜇g IgG with Alexa Fluor® 555
GlyCLICK Fluorophore 555 2 mg
Site-specific conjugation of 2 mg IgG with Alexa Fluor® 555
GlyCLICK® Fluorophore 647 250 ug
Site-specific conjugation of 250 𝜇g IgG with Alexa Fluor® 647
GlyCLICK® Fluorophore 647 2 mg
Site-specific conjugation of 2 mg IgG with Alexa Fluor® 647
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Product Information
Unit Defintion
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.
Contents
Components in GlyCLICK Fluorophore:
- GlycINATOR Immobilized
- UDP-GalNAz
- GalT(Y289L)
- sDIBO (Alexa Fluor 488, 555 or 647)
- All buffers needed
- Concentration and desalting columns
Storage
GlyCLICK Fluorophore 488: Components should be stored at different temperatures.
GlyCLICK Fluorophore 555: Components should be stored at +4-8°C.
GlyCLICK Fluorophore 647: Components should be stored at +4-8°C.
Resources
GlyCLICK® for Generation of Site-specific Conjugates for Fluorescent Imaging
Download Scientific Poster
Applications
Supports high-quality immunofluorescence imaging with consistent signal intensity and reliable quantification through defined antibody labeling.
Higher-resolution flow cytometry with improved signal intensity and a 10-fold increase in separation index using site-specific antibody labeling.
FAQ and Support
Why do I need to use an alkyne-functionalized label with GlyCLICK?
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.
What fluorescent labels can I use with the GlyCLICK technology?
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.
What is the conjugation efficacy using GlyCLICK?
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.
How do I perform quality control of the conjugation process?
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.
Legal & Disclaimers
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Genovis products are intended for research use only. They are not intended to be used for therapeutic or diagnostic purposes in humans or animals. All goods and services are sold subject to Genovis’ General Terms and Conditions of Sale.
SiteClick™ included in GlyCLICK®
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com. The trademark SiteClick™ is the property of Life Technologies Corporation.
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