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S-9011
Protein-Oligo Conjugation Kit


Protein-Oligo Conjugation Kit

The Protein-Oligo Conjugation Kit is designed to easily and efficiently conjugate two separate protein-oligo conjugates. This kit is flexible so that researchers with little or no conjugation experience can make their own custom protein-oligo conjugates to suit their needs. It includes all of the necessary components - including S-HyNic and S-4FB - and protocols for easy and specific crosslinking of any protein with any oligo up to 100 base pairs in length.

Workflow Diagram
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Chromalink™ technology is an innovative, catalyzed, UV-traceable, heterobifunctional linker technology that offers greater efficiency and yield in a considerably simpler method.

  • S-HyNic (succinimidyl-6-hydrazino-nicotinamide) linker is conjugated to the protein through primary amines (-NH2) on the amino acid, lysine, or on the N-terminus.
  • S-4FB (succinimidyl-4-formylbenzamide) linker is conjugated to the oligo.
  • HyNic-modified protein is incubated with 4FB-modified oligo in a catalyzed conjugation.

The result is two biomolecules conjugated through a UV-traceable, stable bond (bis-arylhydrazone) with measurable absorbance at 354 nm.

The TriLink bioconjugation method, with the addition of the TurboLink™ catalyst, guarantees >95% conversion of protein to conjugate when more than 4 molar equivalents of oligo are added. High conversion rates, coupled with the unique UV traceable bond formed during crosslinking, allows for easy purification and identification of the conjugate from the excess oligo using size exclusion purification methods such as FPLC or diafiltration. Each kit contains the material to synthesize two conjugation reactions for any immunoPCR or hybridization-type assay in just under 4 hours each, yielding between 40–60% conjugate after purification.

 

Protein-Oligo Conjugation Kit Advantages

  • Catalyzed conjugation – Faster kinetics for greater efficiency and yields
  • Quantifiable – Using a UV signature wavelength and simple UV scan
  • Stable – 10 times more stable than any other conjugation linker
  • Specificity – Two-linker method avoids homoconjugate formation

 

Additional Information

Citations

  1. Habtemichael EN, Brewer PD, Romenskaia I, Mastick CC. Kinetic evidence that Glut4 follows different endocytic pathways than the receptors for transferrin and alpha-2-macroglobulin. J Biol Chem 2011; 10.1074/jbc.M111.217935. Link 
  2. Grotzky A, Manaka Y, Kojima T, Walde P. Preparation of Catalytically Active, Covalent α-Polylysine−Enzyme Conjugates via UV/Vis-Quantifiable Bis-aryl Hydrazone Bond Formation. Biomacromolecules. 2011:12(1):134-44. Epub 2010 Dec 20. Link
  3. Horikawa S, Bedi D, Li S, Shen W, Huang S, Chen I. Effects of Surface Functionalization on the Surface Phage Coverage and the Subsequent Performance of Phage-Immobilized Magnetoelastic Biosensors. Biosensors and Bioelectronics 2011;26(5):2361-7. Epub 2010 Oct 14. Link 
  4. He J, Wang Y, Dou S, Liu X, Zhang S, Liu G, and Hnatowich D. Affinity Enhancement Pretargeting: Synthesis and Testing of a 99mTc-Labeled Bivalent MORF. 2010 Molecular Pharmaceutics Vol. 7, No. 4, 1118–1124. Link
  5. Shin IS, Maeng JS, Jang BS, You E, Cheng K, Li KCP, Wood B, Carrasquillo JA, Danthi SN, Paik CH. Tc-labeling of Peptidomimetic Antagonist to Selectively Target v 3 Receptor-Positive Tumor: Comparison of PDA and EDDA as co-Ligands. Current radiopharmaceuticals 2010;3(1):1. Link 
  6. Kubler-Kielb J, Majadly F, Biesova Z, Mocca CP, Guo C, Nussenzweig R, Nussenzweig V, Mishra S, Wu Y, Miller LH. A bicomponent Plasmodium falciparum investigational vaccine composed of protein-peptide conjugates. Proceedings of the National Academy of Sciences 2010;107(3):1172. Link 
  7. Chawla K, Ham HO, Nguyen T, Messersmith PB. Molecular resurfacing of cartilage with proteoglycan 4. Acta Biomaterialia 2010;6(9):3388-3394. Link  
  8. Iqbal M, Gleeson MA, Spaugh B, Tybor F, Gunn WG, Hochberg M, Baehr-Jones T, Bailey RC, Gunn LC. Label-Free Biosensor Arrays Based on Silicon Ring Resonators and High-Speed Optical Scanning Instrumentation. Selected Topics in Quantum Electronics, IEEE Journal of 2010;16(3):654-661. Link
  9. Ruggiero A, Villa CH, Holland JP. Imaging and treating tumor vasculature with targeted radiolabeled carbon nanotubes. International Journal of Nanomedicine 2010;5:783-802. Link 
  10. D'Alessandria C, di Gialleonardo V, Chianelli M, Mather SJ, de Vries EF, Scopinaro F, Dierck RA, Signore A. Synthesis and Optimization of the Labeling Procedure of 99mTc-Hynic-Interleukin-2 for In vivo Imaging of Activated T lymphocytes. Mol Imaging Biol. 2010;12(5): 539–546. Link 
  11. Liu G, Dou S, Chen X, Chen L, Liu X, Rusckowski M, Hnatowich DJ. Adding a clearing agent to pretargeting does not lower the tumor accumulation of the effector as predicted. Cancer Biother Radiopharm. 2010;25(6):757-62. Link  
  12. Washburn AL, Gunn LC, Bailey RC. Label-free quantitation of a cancer biomarker in complex media using silicon photonic microring resonators. Analytical chemistry 2009;81(22):9499-9506. Link
  13. Johnson LM, Avens HJ, Hansen RR, Sewell HL, Bowman CN. Characterization of the Assaying Methods in Polymerization-Based Amplification of Surface Biomarkers. Australian Journal of Chemistry 2009;62(8):877-884. Link
  14. Hansen RR, Johnson LM, Bowman CN. Visual, base-specific detection of nucleic acid hybridization using polymerization-based amplification. Analytical biochemistry 2009;386(2):285-287. <NO LINK>
  15. Myles W. Gardner MW and Brodbelt JS. Ultraviolet Photodissociation Mass Spectrometry of Bis-aryl Hydrazone Conjugated Peptides. Anal. Chem.2009,81,4864-4872. Link  
  16. Liu G, Cheng D, Dou S, Chen X, Liang M, Pretorius PH, Rusckowski M, Hnatowich DJ. Replacing 99m Tc with 111In Improves MORF/cMORF Pretargeting by Reducing Intestinal Accumulation. Molecular Imaging and Biology 2009;11(5):303-307. Link 
  17. Chen X, Dou S, Liu G, Liu X, Wang Y, Chen L, Rusckowski M, Hnatowich DJ. Synthesis and in Vitro Characterization of a Dendrimer- MORF Conjugate for Amplification Pretargeting. Bioconjugate chemistry 2008;19(8):1518-1525. Link 
  18. Kattah MG, Coller J, Cheung RK, Oshidary N, Utz PJ. HIT: a versatile proteomics platform for multianalyte phenotyping of cytokines, intracellular proteins and surface molecules. Nature medicine 2008;14(11):1284-1289. Link 
  19. Sharma S, Dominguez AL, Manrique SZ, Cavallo F, Sakaguchi S, Lustgarten J. Systemic Targeting of CpG-ODN to the Tumor Microenvironment with Anti–neu-CpG Hybrid Molecule and T Regulatory Cell Depletion Induces Memory Responses in BALB-neuT Tolerant Mice. Cancer research 2008;68(18):7530. Link 
  20. Chaturvedi A, Dorward D, Pierce SK. The B cell receptor governs the subcellular location of Toll-like receptor 9 leading to hyperresponses to DNA-containing antigens. Immunity 2008;28(6):799-809. Link 
  21. Qi H, Cannons JL, Klauschen F, Schwartzberg PL, Germain RN. SAP-controlled T–B cell interactions underlie germinal centre formation. Nature 2008;455(7214):764-769. Link  
  22. Liu G, Dou S, Rusckowski M, Hnatowich DJ. An experimental and theoretical evaluation of the influence of pretargeting antibody on the tumor accumulation of effector. Molecular cancer therapeutics 2008;7(5):1025. Link 
  23. Liu G, Dou S, Pretorius PH, Liu X, Rusckowski M, Hnatowich DJ. Pretargeting CWR22 prostate tumor in mice with MORF-B72.3 antibody and radiolabeled cMORF. Eur J Nucl Med Mol Imaging. 2008 Feb;35(2):272-80. Link 
  24. Walls ZF, Gambhir SS. BRET-based method for detection of specific RNA species. Bioconjugate chemistry 2008;19(1):178-184. Link 
  25. Liu G, Dou S, He J, Liu X, Rusckowski M, Hnatowich DJ. Predicting the biodistribution of radiolabeled cMORF effector in MORF-pretargeted mice. European journal of nuclear medicine and molecular imaging 2007;34(2):237-246. Link 
  26. Liu G, Dou S, Yin D, Squires S, Liu X, Wang Y, Rusckowski M, Hnatowich DJ. A novel pretargeting method for measuring antibody internalization in tumor cells. Cancer biotherapy & radiopharmaceuticals 2007;22(1):33-39. Link 
  27. Buhl A, Metzger JH, Heegaard NHH, von Landenberg P, Fleck M, Luppa PB. Novel biosensor-based analytic device for the detection of anti-double-stranded DNA antibodies. Clinical chemistry 2007;53(2):334. Link  
  28. Bailey RC, Kwong GA, Radu CG, Witte ON, Heath JR. DNA-encoded antibody libraries: a unified platform for multiplexed cell sorting and detection of genes and proteins. J. Am. Chem. Soc 2007;129(7):1959-1967. Link 
  29. He J, Liu G, Dou S, Gupta S, Rusckowski M, Hnatowich D. An improved method for covalently conjugating morpholino oligomers to antitumor antibodies. Bioconjugate Chem 2007;18(3):983-988. Link  

 

Not for resale without express written permission. Not for use in humans. No license under any patent or patent pending is granted or implied by the purchase of any TriLink product. TriLink does not warrant that the use or sale of the products delivered hereunder will not infringe the claims of any United States or other patents or patents pending covering the use of the product alone or in combination with other products or in the operation of any process. All and any use of TriLink product is the purchaser’s sole responsibility.




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   Kit - 2 Conjugations ($473.00)


 



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