Antibody Biotinylation Kit

Albert Einstein College of Medicine
Microbiology & Immunology
PhD Candidate

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Scientific

Product Name:

EZ-Link™ Sulfo-NHS-LC-Biotinylation Kit

Catalog Number:

21435

We used this kit to biotynilate an antibody to be used in FACS experiments to detect cells that are expressing HIV gp120. The antibody we wanted to use was a broadly neutralizing antibody that was isolated from a patient and therefore there was no commercially available form for flow cytometry.

Experimental Design and Results Summary

Application

Antibody Biotinylation

Starting Material

Purified antibody of interest (1-10mg)

Protocol Overview

Calculate the amount of biotin reagent to add to the reaction based on the amount of antibody you are starting with (follow calculation example in the protocol). Add the appropriate amount of biotin reagent to the antibody solution. Incubate on ice for 2h. Perform a buffer exchange using the Zeba Spin Column provided in the kit.

Tips

Follow the protocol of the kit and double check calculations

Results Summary

The resulting Biotinylated antibody was used to stain cells expressing HIV gp120 following by staining with streptavidin-PE. Staining worked well and the antibody was stable for at least a year after biotinylation.

DOI or PMID #

N/A

Additional Notes

None

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Summary

The Good

Works well for antibody biotinylation

The Bad

Not direct labeling. For FACS need to follow with Streptaviding-dye

The Bottom Line

Good kit to use if you want to label a large amount of antibody (1-10mg).

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