Calculator
Dye to Nucleic Acid Conjugation
Protein to nucleic acid conjugation is a chemical strategy to couple a protein molecule to an oligonucleotide molecule. The attachment of protein molecule, such as antibody and enzyme, provides traceability and detectability for the oligonucleotide through antibody-antigen recognition or enzymatic reactions that can produce chromogenic or fluorescent products.
The conjugation reactions involved in protein- oligonucleotide cross-linking are similar as the methods used to form protein-protein conjugates. The only requirement is that the oligonucleotide molecule to be modified should contain one or more suitable reactive groups, such as amines or sulfhydryles, which could be successfully introduced in the oligonucleotide strand during the solid-phase synthesis process. Subsequently, bifunctional cross-linkers, like SMCC, can be used to couple protein molecules to the modified oligonucleotide using the same basic principles effective in protein-protein conjugations.
AAT Bioquest has developed an optimized and robust Buccutite™ crosslinking technique. Buccutite™ crosslinking technology provides a simplistic and efficient approach for conjugating proteins to nucleic acids. It can be completed in half the time that traditional SMCC cross-linker needs and with less stringent parameters yielding highly stable and easy to handle conjugated proteins.
The conjugation reactions involved in protein- oligonucleotide cross-linking are similar as the methods used to form protein-protein conjugates. The only requirement is that the oligonucleotide molecule to be modified should contain one or more suitable reactive groups, such as amines or sulfhydryles, which could be successfully introduced in the oligonucleotide strand during the solid-phase synthesis process. Subsequently, bifunctional cross-linkers, like SMCC, can be used to couple protein molecules to the modified oligonucleotide using the same basic principles effective in protein-protein conjugations.
AAT Bioquest has developed an optimized and robust Buccutite™ crosslinking technique. Buccutite™ crosslinking technology provides a simplistic and efficient approach for conjugating proteins to nucleic acids. It can be completed in half the time that traditional SMCC cross-linker needs and with less stringent parameters yielding highly stable and easy to handle conjugated proteins.
Table 1. Buccutite™ vs SMCC Conjugation At-A-Glance
| Conjugation Method ▲ ▼ | Buccutite™ Kits ▲ ▼ | SMCC Conjugation ▲ ▼ |
| Minimum antibody concentration | 100 µg/mL | 0.5 to 5 mg/mL |
| Labeling Chemistry | Uses two proprietary linkers, Buccutite™ MTA and Buccutite™ FOL, to facilitate protein-protein conjugation. | Uses a heterobifunctional crosslinker to facilitate protein-protein conjugation. Labeling protocol is tedioius and requries existing knowledge of bioconjugation chemistry. |
| Available labels | PE, APC, Tandem Dyes, HRP, Poly-HRP and AP. Cat No 1315 can be used to conjugate any two proteins. | User is responsible for supplying proteins, method suitable for any protein-protein conjugation |
| Type of bond between label and antibody | Covalent | Covalent |
| Compatible with BSA or other stabilizers? | No | No |
| Requires purification? | No | Yes |
| Time required for conjugation | 2 hrs | 4 hrs or more |
| Hands-on-time | ~15 minutes | 2 hrs |
| Conjugate yield | 100 % | ~30% |
| Batch-to-batch variation | Minimal | High |