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MAGICLINK™ Site Specific Kits provide optimized reagents for labeling antibodies. The most important features of the kits are ultra-fast reaction speed, high conjugation efficiency, and the most stable linkage between antibody and oligonucleotide on the market to date. Labeling reactions chemiselectively target disulfide bonds for maximum labeling efficiency. The kits provide optimized reagents for conjugating amine modified oligonucleotide to antibodies or other proteins with disulfide bonds. Each 100 μg kit reaction is sufficient for labeling 50-200 μg of antibody in 100 μL reaction volumes, and 500 μg kit for 500 - 1000 μg antibody. The MAGIC Site-Specific linker, provided as a controlled excess reagent, reacts with the antibody under mild alkaline pH conditions. Similarly, the LINK ester will react with the amine-modified oligonucleotide under the same condition. Prior to coupling, LINK-oligonucleotide and MAGIC-antibody are desalted with centrifugal concentrators. The MAGIC antibody then reacts instantly with LINK-oligonucleotide to get the oligonucleotide-antibody conjugates.
Additionally, the kits include LINK-650 dye to determine the MAGIC number on activated antibody. These kits can be used to conjugate antibodies to single-stranded oligos that are 10-120 bases long or to double-stranded oligos that are up to 80 bases long. The oligos is amine-modified one which can be done in a regular Oligo synthesis lab. Current antibody labeling methods, such as amine-NHS ester and converting amine to thiol and then use maleimide chemistry, will produce random labels on the antibody. In some cases, labeling in Fab areas could block antigen binding. With site-specific cysteine/disulfide chemistry, these linkers are inserted to disulfice bond location to form thiol-bridgend do not affect antibody-antigen binding (Fig.1). It will help increase the sensitivity of bio assay such as ELISA, Western Blot, etc. 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 user guide.
MagicLink Oligo antibody Labeling Kits are used to conjugate oligo with primary antibody, secondary antibodies, proteins, etc. with available cysteine and disulfides for the bioassays below. Oligo-antibody applications include:
Oligozymes application: dot blots, Southern/Northern blots, In situ, and solution hybridization/capture