Buccutite™ Bioconjugation Technology
Rapid Protein Crosslinking Method for Labeling and Modifying Antibodies
Crosslinking is a technique that chemically joins two or more proteins together, such as an antibody and an enzyme, by a covalent bond. Crosslinking reagents consist of reactive moieties that modify and attach to specific functional groups on proteins. A common approach for conjugating two biomolecules utilizes a small, heterobifunctional crosslinker called succinimidyl 4-(N-maleimidomethyl) cyclohexane1-carboxylate (SMCC). SMCC crosslinkers contain amine-reactive NHS ester moieties and thiol-reactive maleimide moieties to crosslink proteins. However, a drawback to this approach is the many experimental conditions that must be monitored to perform a successful conjugation. Difficulties associated with SMCC conjugations are maintenance of a narrow pH range, self-polymerization, and hydrolytic degradation of reactive moieties. To address these concerns, AAT Bioquest has developed an optimized and robust Buccutite™ crosslinking technique. Buccutite™ crosslinking technology provides a simplistic and efficient approach for conjugating proteins. It can be completed in half the time and with less stringent parameters yielding highly stable and easy to handle conjugated proteins.
Buccutite™ Crosslinking Principles
Conjugate Proteins In 2 Simple Mixing Steps. No catalyst required
Buccutite™ vs SMCC
Total Operation Time:
- Buccutite™: 2 Hours
- SMCC Chemistry: 4-8 Hours
Minimum Sample Concentration
- Buccutite™ Chemistry: ≥0.5 µg/µL
- SMCC Chemistry: 0.5 - 5.0 µg/µL
Optimal Conjugation pH:
- Buccutite™ Chemistry: 5 - 9
- SMCC Chemistry: 7.0 - 7.5
Conjugation Yield
- Buccutite™ Chemistry: 50-60 %
- SMCC Chemistry: ≤ 30 %
Advantages
Buccutite™ conjugation can be run in a broad pH range of 5-9 and temperature.
Fast: Buccutite™ conjugation can be completed in 1-2 hours under extremely mild conjugation condutions. No purification step required with 25μg kits.
Robust: Buccutite™ conjugation can be run in a broad pH range of 5-9 and temperature.
Highly Stable: Buccutite™ linker-activated macromolecules are very stable, can be stored at 4ºC for more than 24 months.
High Yield: Buccutite™ conjugation gives much higher yield than other existing methods under the same conditions. Eliminating homo-crosslinking of proteins.
Tubulins were imaged with RPE- goat-anti-mouse IgG conjugate in HeLa cells. Tubulins were stained using mouse anti-α-tubulin antibody, and visualized with red fluorescent RPE-goat-antimouse IgG conjugate prepared by Buccutite™ crosslinking technology as described above.
AAT Bioquest’s Custom Bioconjugation Services
AAT Bioquest offers same-day custom conjugation of proteins or antibodies with a wide array of labels, such as biotin, HRP and over 20 fluorophores. Our services are designed to produce high-quality results with a fast turnaround. We take pride in supplying scientists with flexible tools that help expand their research needs.
Features of AAT Bioquest’s custom bioconjugation services include:
- Guaranteed quality with 95% purity rating
- Affordable price tiers that work with your budget
- Scalable service with minimum order of 50 µg
- Same day order fulfilment means your conjugate ships the day you order it
*NOTE: APC and PE custom conjugation services require 3 working days You may supply your own protein or antibody or chose from 3000+ monoclonal and polyclonal antibodies in our catalog. Contact us for a quote today, and let us put our expertise to work for you!
Product Ordering Information For Buccutite™ Labelling Kits
Cat. # | Product Name | Abs/Em (nm) |
5503 | Buccutite™ Peroxidase (HRP) Antibody Conjugation Kit *Optimized for Labeling 100 µg Protein* | N/A |
1313 | Buccutite™ Rapid APC Antibody Labeling Kit *Microscale Optimized for Labeling 2w5 μg Antibody Per Reaction* | 651/662 |
1311 | Buccutite™ Rapid APC Antibody Labeling Kit *Microscale Optimized for Labeling 100 μg Antibody Per Reaction* | 651/662 |
1322 | Buccutite™ Rapid APC-Cy5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 µg Antibody Per Reaction* | 651/670 |
1350 | Buccutite™ Rapid APC-Cy5.5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 µg Antibody Per Reaction* | 651/700 |
1320 | Buccutite™ Rapid APC-Cy5.5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 µg Antibody Per Reaction* | 651/700 |
1351 | Buccutite™ Rapid APC-Cy7 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 µg Antibody Per Reaction* | 651/780 |
1321 | Buccutite™ Rapid APC-Cy7 Tandem Antwibody Labeling Kit *Microscale Optimized for Labeling 100 µg Antibody Per Reaction* | 651/780 |
1347 | Buccutite™ Rapid APC-iFluor® 700 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 µg Antibody Per Reaction* | 651/713 |
1319 | Buccutite™ Rapid APC-iFluor® 700 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 µg Antibody Per Reaction* | 651/713 |
1312 | Buccutite™ Rapid PE Antibody Labeling Kit *Microscale Optimized for Labeling 25 μg Antibody Per Reaction* | 565/575 |
1310 | Buccutite™ Rapid PE Antibody Labeling Kit *Microscale Optimized for Labeling 100 μg Antibody Per Reaction* | 565/575 |
1340 | Buccutite™ Rapid PE-Cy5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 µg Antibody Per Reaction* | 565/670 |
1341 | Buccutite™ Rapid PE-Cy5.5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 µg Antibody Per Reaction* | 565/700 |
1316 | Buccutite™ Rapid PE-Cy5.5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 µg Antibody Per Reaction* | 565/700 |
1342 | Buccutite™ Rapid PE-Cy7 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 µg Antibody Per Reaction* | 565/700 |
1317 | Buccutite™ Rapid PE-Cy7 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 µg Antibody Per Reaction* | 565/780 |
1353 | Buccutite™ Rapid PerCP Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 µg Antibody Per Reaction* | 482/677 |
1325 | Buccutite™ Rapid PerCP Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 µg Antibody Per Reaction* | 482/677 |
1343 | Buccutite™ Rapid PE-Texas Red Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 µg Antibody Per Reaction* | 565/600 |
1318 | Buccutite™ Rapid PE-Texas Red Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 µg Antibody Per Reaction* | 565/600 |
1315 | Buccutite™ Rapid Protein Crosslinking Kit *Microscale Optimized for Crosslinking 100 μg Antibody Per Reaction* | N/A |
Original created on September 12, 2017, last updated on September 12, 2017
Tagged under: Antibody & Protein Labeling