Thermo Scientific
EZ-Link™ Sulfo-NHS-LC-Biotinylation Kit
21435
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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.
Antibody Biotinylation
Purified antibody of interest (1-10mg)
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.
Follow the protocol of the kit and double check calculations
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.
N/A
None
Works well for antibody biotinylation
Not direct labeling. For FACS need to follow with Streptaviding-dye
Good kit to use if you want to label a large amount of antibody (1-10mg).
Product Name: EZ-Link Sulfo-NHS-LC-Biotinylation Kit, 10-rxn kitSupplier Page