GlyCLICK®
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GlyCLICK® Azide Activation
Site-specific conjugation of IgG with azide-alkyne click chemistry
GlyCLICK is a three-step conjugation technology for site-specific and quantitative conjugation of IgG from several species and subclasses.
GlyCLICK Azide Activation contains all reagents needed to azide-activate 250 μg, 2 mg or 10 mg IgG for site-specific custom conjugation using any alkyne-reactive label of choice. Are you in need of azide activation of larger amounts of IgG? Contact us.
Why GlyCLICK® Azide Activation?
- Site-specific azide activation of IgG with a degree of labeling (DOL) of 2
- Versatile and consistent custom conjugation of IgG
- Fc-specific labeling preserves antigen-binding capacity
Available Products
GlyCLICK® Azide Activation 250 ug
Site-specific conjugation of 250 𝜇g IgG with azide-alkyne click chemistry
GlyCLICK® Azide Activation 2 mg
Site-specific conjugation of 2 mg IgG with azide-alkyne click chemistry
GlyCLICK® Azide Activation 10 mg
Site-specific conjugation of 10 mg IgG with azide-alkyne click chemistry
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Product Information
Unit Defintion
GlyCLICK Azide Activation contains sufficient material for azide activation of 250 μg, 2 mg or 10 mg IgG. For larger amounts or evaluation of the technology, please contact us.
Contents
Components in GlyCLICK Azide Activation:
- GlycINATOR Immobilized
- UDP-GalNAz
- GalT(Y289L)
- All buffers needed
- Concentration and desalting columns
Storage
GlyCLICK Azide Activation 250 μg / 2 mg: Components should be stored at +4-8°C upon arrival.
GlyCLICK Azide Activation 10 mg: Components should be stored at different temperatures upon arrival.
Resources
GlyCLICK®: From Kit to Clinic
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FAQ and Support
What is the smallest amount of IgG that I can process using the 250 ug kit?
The GlyCLICK 250 ug kit is optimized for 250 ug of IgG as a starting material. We do not recommend using less than 100 ug of starting material in a minimum of 100µL to achieve proper end-over-end mixing on the GlycINATOR columns. Please contact support@genovis.com for assistance if you want to process smaller amounts then 100µg.
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.
For how long can I store azide activated antibodies?
The stability of azide activated material varies depending on the antibody and if stabilizing agents have been added. In our labs we have stored azide activated antibodies in TBS buffer without any additives at +4°C for over two years. The azide activation was still OK, and the click reaction generated antibodies with DOL=2. If azide is added as a preservative during long-term storage, it must be removed prior to click reaction.
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 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.
Legal & Disclaimers
All rights reserved. Genovis products may be covered by one or more patents, trademarks and copyrights owned by Genovis AB or licensed from third parties. For more information about commercial rights, please contact the Genovis team at licensing@genovis.com.
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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