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Biotin-dPEG®₄-hydrazide (QBD-10219)

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Description

Biotin-dPEG®4-hydrazide, product number QBD-10219, is a hydrophilic biotinylation reagent designed for labeling the carbohydrate coats of glycoproteins with biotin. The amphiphilic spacer provides excellent water solubility for poorly water-soluble biotin. Moreover, the dPEG®4 spacer is longer than the hydrophobic aminocaproic acid (LC) spacer used in biotin-LC-hydrazide, so it provides somewhat better access to the biotin binding pockets of avidin and streptavidin. Additionally, the dPEG®4 spacer reduces or eliminates non-specific binding in many applications.

Biotin-dPEG®4-hydrazide, QBD-10219, is a carbonyl-reactive compound that forms semi-permanent hydrazone bonds with aldehydes, ketones, and carboxylic acid groups. Although there are numerous natural small molecules containing aldehydes and ketones, macromolecules of biological importance (peptides, proteins, and nucleic acids) rarely contain them. However, glycosylated proteins such as antibodies frequently contain carbohydrate residues known as reducing sugars that can be oxidized with sodium periodate, neuraminidase, or galactose oxidase to form aldehydes.

The aldehydes thus formed then can be reacted with Biotin-dPEG®4-hydrazide at pH 5 – 7, labeling the protein with biotin through the formation of a Schiff base. Aniline or para-phenylenediamine catalyzes the reaction, forming the Schiff base quickly. For additional bond stability, sodium cyanoborohydride can be used to reduce the Schiff base to a secondary amine. Via carbodiimide chemistry (for example, 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide, also known as EDC), Biotin-dPEG®4-hydrazide reacts with carboxylic acid groups forming a stable bond.

The amphiphilic dPEG®4 spacer in Biotin-dPEG®4-hydrazide provides the ability to use Biotin-dPEG®4-hydrazide directly in water or aqueous buffer, making it more biocompatible than LC-biotin. Moreover, QBD-10219 will not trigger protein or peptide aggregation or cause non-specific binding due to hydrophobicity issues from the linker, both of which can occur with traditional biotin linkers like LC-biotin.

Biotin-dPEG®4-hydrazide is highly useful for installing a biotin label onto glycoproteins. It is also used in cell labeling to label glycoproteins on cell surfaces. Biotinylated glycoproteins can be identified using streptavidin-horseradish peroxidase (HRP) conjugates, because of the high affinity between biotin and streptavidin. Biotinylated secondary antibodies are used frequently with avidin or streptavidin in plate-based assays such as ELISA and western blots. This reagent, coupled with avidin that has been modified to have reduced biotin-binding affinity, is for affinity purification of molecules.

Specifications

Unit Size50 mg, 1000 mg
Molecular Weight505.63; single compound
Chemical formulaC₂₁H₃₉N₅O₇S
CAS756525-97-0
Purity> 98%
SpacersdPEG® Spacer is 18 atoms and 20.6 Å
ShippingAmbient
Typical solubility properties (for additional information contact Customer Support)DMAC, DMSO or water; it is not soluble in acetonitrile.
Storage and handling-20°C; Always let come to room temperature before opening; be careful to limit exposure to moisture and restore under an inert atmosphere; stock solutions can be prepared with dry solvent and kept for several days (freeze when not in use). dPEG® pegylation compounds are generally hygroscopic and should be treated as such. This will be less noticeable with liquids, but the solids will become tacky and difficult to manipulate, if care is not taken to minimize air exposure.

References

Greg T. Hermanson, Bioconjugate Techniques, 2nd Edition, Elsevier Inc., Burlington, MA 01803, April, 2008 (ISBN-13: 978-0-12-370501-3; ISBN-10: 0-12-370501-0); See pp. 733-736 for a general discussion of the chemistry and a general protocol for labeling of glycoprotein is given on p. 736.

Greg T. Hermanson, Bioconjugate Techniques, 3rd Edition, Elsevier, Waltham, MA 02451, 2013, ISBN 978-0-12-382239-0; See Chapter 18, Discrete PEG Reagents, pp. 787-821, for a full overview of the dPEG® products.

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Electrochemically Directed Modification of ITO Electrodes and Its Feasibility for the Immunosensor Development. Daegeun Yu and Kyuwon Kim. Bull Korean Chem. Soc.2009, 30 (4) pp 955-958. February 23, 2009.

WSS25 Inhibits Growth of Xenografted Hepatocellular Cancer Cells in Nude Mice by Disrupting Angiogenesis via Blocking Bone Morphogenetic Protein (BMP)/Smad/Id1 Signaling. Hong Qiu, Bo Yang, Zhi-Chao Pei, Zhang Zhang and Kan Ding. JBC. 2010, 285 (42) pp 32638–32646. October 15, 2010. DOI: 10.1074/jbc.M110.105544.

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Simultaneous Monitoring of Presynaptic Transmitter Release and Postsynaptic Receptor Trafficking Reveals an Enhancement of Presynaptic Activity in Metabotropic Glutamate Receptor-Mediated Long-Term Depression. Wei Xu, Yiu Chung Tse, Frederick A. Dobie, Michel Baudry, Ann Marie Craig, Tak Pan Wong, and Yu Tian Wang. The Journal of Neuroscience. 2013, 33 (13) pp 5867-5877. March 27, 2013. DOI:10.1523/JNEUROSCI.1508-12.2013.

Three Recombinant Engineered Antibodies against Recombinant Tags with High Affinity and Specificity. Hongyu Zhao, Ao Shen, Yang K. Xiang, and David P. Corey. PLoS ONE. 2016, 11 (3) e0150125. March 4, 2016. DOI: 10.1371/journal.pone.0150125.

The carbohydrate-linked phosphorylcholine of the parasitic nematode product ES-62 modulates complement activation. Umul Kulthum Ahmed, N. Claire Maller, Asif J. Iqbal, Lamyaa Al-Riyami, William Harnett, and John G. Raynes. The Journal of Biological Chemistry. 2016, April 4, 2016. DOI: 10.1074/jbc.M115.702746.

A Facile Approach to Functionalize Cell Membrane-Coated Nanoparticles. Hao Zhou, Zhiyuan Fan, Pelin K. Lemons, and Hao Cheng. Theranostics. 2016, 6 (7) pp 1012-1022. April 28, 2016. DOI: 10.7150/thno.15095.

Epidermal-specific deletion of CD44 reveals a function in keratinocytes in response to mechanical stress. M Shatirishvili, AS Burk, CM Franz, G Pace, T Kastilan, K Breuhahn, E Hinterseer, A Dierich, L Bakiri, EF Wagner, H Ponta, TN Hartmann, M Tanaka, and V Orian-Rousseau. Cell Death and Disease. 2016, 7 (e2461). November 10, 2016. DOI: 10.1038/cddis.2016.342.

Antibody neutralization of CXCL10 in vivo is dependent on binding free and not endothelial bound chemokine: Implications for the design of a new generation of anti-chemokine therapeutic antibodies. Pauline Bonvin, Franck Gueneau, Vanessa Buatois, Maud Charreton-Galby, Stanley Lasch, Marie Messmer, Urs Christen, Andrew D Luster, Zoe Johnson, Walter Ferlin, Marie Kosco-Vilbois, Amanda Proudfoot, and Nicolas Fischer. The Journal of Biological Chemistry. 2017, 292 (10), pp 4185-4197. January 30, 2017. DOI: 10.1074/jbc.M116.745877.

The Migrastatin Family:  Discovery of Potent Cell Migration Inhibitors by Chemical Synthesis. Christoph Gaul, Jón T. Njardarson, Dandan Shan, David C. Dorn, Kai-Da Wu, William P. Tong, Xin-Yun Huang, Malcolm A. S. Moore, and Samuel J. Danishefsky. Journal of the American Chemical Society. 2004, 126 (36) pp 11326-11337. DOI: 10.1021/ja048779q.

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Exploitation of SPR to Investigate the Importance of Glycan Chains in the Interaction between Lactoferrin and Bacteria. Noelle O’Riordan, Michelle Kilcoyne, Lokesh Joshi, and Rita M. Hickey. Senors. 2017, 17 (7) p 115. June 27, 2017. doi:10.3390/s17071515.

Producing A Peptide For Use In A Blood Biosensor For Injury Detection. Errek Manh Trung Pham. Youngstown State University Theses & Dissertations Center. 2020. December 2020. ohiolink.edu/1607519672342672

Applicable patents and legal notices are available at legal notices.

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