Products

MagicLink™ Oligo Antibody Conjugation Kit

Oligonucleotide can be conjugated to antibody/protein by thiol-maleimide and hydrazone coupling

approaches. The thiol-maleimide approach is commonly performed with a bifunctional linker (such as

SMCC). However, SMCC-modified protein is extremely unstable and often self-reactive since it often

contains both amine and thiol groups that cause a significant amount of homo-crosslinking. The conjugation

linkage is exchangeable when other thiol groups are present. In the hydrazonecoupling approach, the

conjugation reaction is based on a reaction between hydrazine and aldehyde. This method is low yielding

although a catalyst is used. The size of the molecule and inaccessibility of the functional group contribute to

low yield. In addition, linkage stability is an issue.

A typical flow chart of oligo-antibody conjugation by MagicLink technology

BroadPharm's MagicLink™ technology solves all the problems by combining PEG and proprietary

crosslinking chemistry. MagicLink™ chemistry and kit do not need TCEP, DTT, or catalysts at all!

First, antibody reacts with LINK NHS ester to introduce LINK into the antibody. Also, amine modified

oligonucleotide reacts with MAGIC NHS ester to introduce MAGIC into the oligonucleotide structure. Second,

after purification of both reactive partners, they instantly react with each other yielding an antibody-

oligonucleotide conjugate with a stable ring linkage.

Features;

· Novelty: 3rd generation protein crosslink technology

· Yield: conjugation efficiency around 95-100%

· Rate: instant reaction; the whole process less than 1 hour

· Linkage stability: most stable covalent bonded conjugates on the market

· Consistence: good reproducibility from batch to batch

· Flexible: use your own HRP and antibody at your desired ratio

· Shelf life: long, stable freeze dried ship at ambient temperature

· Antibody selection: any proteins/antibody, antibody fragment

· Oligo selection: single strand oligos of 10 120 bases, and double strand oligos up to 80 base pairs; Linking

  chemistry works at both 5 and 3 end

· Post-conjugation clean-up step not necessary

· Positive control antibody and oligo included to enable confirmation of protocol success

The kit can also be used to conjugate oligonucleotides to alkaline phosphatase (AP), horseradish

peroxidase (HRP), bovine serum albumin (BSA), soybean peroxidase (SBP), penicillinase (PNC),

streptavidin and its conjugates, alpha-D-galactosidase, glucose-t-phosphate dehydrogenase, etc.

Such oligonucleotide-enzyme conjugates ("oligozymes") can be used in a variety of hybridization and

detection formats, including dot blots, Southern/Northern blots, in situ and solution hybridization/capture

schemes.

SDS-PAGE analysis of antibody conjugates by 4-12% bis- tris gel - the control oligo was conjugated to goat

lgG at different oligo/antibody ratio.

Applications;

MagicLink Oligo antibody Labeling Kits are used to conjugate oligo with primary antibody, secondary

antibodies, enzymes, etc. for the bioassays below.

Oligo-antibody application:

· Immuno-Polymerase Chain Reaction (immuno-PCR)

· Proximity Ligation Assay (PLA)

· Electrochemical Proximity Assay (ECPA)

· Multiplexed Assay Development

· Proximity extension assay

· 3DNA technology in lateral flow

· 3DNA technology in cellular targeting

· Single cell CITE-seq experiments

· DNA-PAINT imaging

· Protein arrays

Oligozymes application:

· Dot blots,

· Southern/Northern blots,

· In situ, and solution hybridization/capture

Consists of;

· MAGIC NHS (MW ~900)

· LINK NHS (MW ~900)

· Oligo control,

· Antibody control

· Protein concentrator

· Reaction buffer

  Instruction sheet (PDF)