Oligo-antibody conjugates have been widely used in research to serve as oligo-delivery tools or detection methods. RayBio^® Oligonucleotide Conjugation Kit provides a simple and highly efficient method to generate oligo-antibody conjugates with less than 20 minutes of on-hand time. This kit can be used to generate the conjugates of different oligo:antibody ratios and achieve up to 90% or higher conjugation efficiency (percentage of antibody conjugated with at least one oligo).

The oligos should be synthesized to include a terminal amine group at either 5’ or 3’ end and purified by HPLC to remove any impurities that may affect the conjugation. The antibody intended for conjugation should also be purified and must be free from BSA, Tris-Glycine, or any other amine-containing impurities, as these can substantially impact the conjugation process. Each reaction is designed to conjugate 100 μg of antibody and a control. This kit contains enough reagents to run 2 reactions or 10 reactions.

As expected for any chemical conjugation reaction, the concentration and buffer formulation of the oligo and the antibody need to fall within certain parameters, as detailed in this booklet. A single-strand oligo (68 nt) and a monoclonal antibody are also provided as positive controls.

• Precision pipettes to deliver 2 μL to 1 mL
• Nuclease-free barrier filter tips fitting the pipettes. Elongated 10 μL tips are recommended to preventcontamination when processing multiple oligos
• Nuclease-free water
• Nuclease-free 1.5-2 mL microcentrifuge tubes
• Terminal amine-modified oligonucleotides for conjugation (HPLC grade)
• Antibodies for conjugation
• Vortex
• Variable-speed benchtop microcentrifuge

1. Calculate oligo:antibody ratio (1:10 is standard)
2. Oligo activation
3. Oligo purification
4. Prepare controls
5. Conjugation
6. Analysis of the conjugate
7. Conjugate purification (if necessary for downstream use)

Upon receipt, store the vial of crosslinker, control antibody, control oligo, and desalting column at 4 ºC and other kit components at room temperature.
Note: once resuspended, the control oligo, crosslinker should be stored at -20ºC, while the control antibody should be stored at 4ºC. Each vial of the Crosslinker Reagent is enough for conjugating 100 μg desired antibody and 30 μg Control Antibody at the same time.