Creative Biolabs offers several high-quality biotin labeling services for our global customers for biomedical research use. Based on a wealth of experience and advanced technology, we are capable of designing and manufacturing both custom and customized antibody conjugation projects in close cooperation with our clients. In particular, our skilled experts enable us to offer quick and optimized conjugate results with excellent quality.
Biotin, also known as vitamin B7, is a small biologically active molecule that acts as a coenzyme in living cells. It can be coupled to oligonucleotides at the terminal and internal position at multiple sites. Biotin labeling (also called biotinylation) is the process of covalently attaching biotin to a protein, nucleic acid, or other molecule, which has been widely used in numerous detection systems especially when the target of interest is expressed at low levels and cannot be detected by labeled antibodies alone. Generally, biotinylation is rapid, and specific and is unlikely to perturb the natural function of the molecule due to the small size of biotin. Most importantly, many biotin molecules can be conjugated to an antibody with extremely high affinity, fast-on-rate, and high specificity, which does not appear in the conjugation with streptavidin and avidin. Through this amplification step, proteins which are expressed at low levels are more likely to be detected.
Fig.1 Biotin maleimide cysteine reaction.1
The biotin tag can be used in the technique of affinity chromatography in conjunction with a column attached to avidin (or streptavidin or neutravidin), the natural ligand for biotin. Tagging the protein with iminobiotin can be utilized to isolate the tagged protein since the biotin analog has significant binding to avidin/streptavidin at alkaline pH but loses affinity when the pH is lowered. As a result, an avidin/streptavidin column can release an iminobiotin-tagged functional protein by reducing the pH (to around pH 4).
Anti-biotin antibodies or avidin/streptavidin-tagged detection techniques such as enzyme reporters or fluorescence probes, which can be employed to detect the protein, may facilitate localization using fluorescence or electron microscopy, ELISA assays, ELISPOT assays, western blots, and other immunoanalytical approaches. It is vital to highlight that utilizing monovalent streptavidin for detection prevents aggregation or aggregation of biotinylated targets.
The non-covalent bond formed by biotin and avidin or streptavidin is nearly as strong as a covalent bond and has a higher binding affinity than most antigen and antibody interactions. Based on this highly strong binding, labeling proteins with biotin is a useful tool for applications such as affinity chromatography, which separates the biotinylated protein from a mixture of other proteins and biochemicals using immobilized avidin or streptavidin. Biotinylation of MHC molecules to generate MHC multimers has also been shown to be an efficient approach for detecting and separating antigen-specific T-cell populations.
Protein biotinylation in living cells is developed to explore protein-protein interactions and proximity. One of the interacting proteins is biotinylated, while the other is being detected using a streptavidin-conjugated detection method. This method is frequently employed in pull-down assays and proximity-dependent labeling procedures.
Having provided protein biotin labeling service for many years, Creative Biolabs is capable of offering a full range of custom biotinylation services for both academic and industrial customers. With experienced scientists, we can label any protein with biotin as our clients require. Our advanced protein biotinylation platform enables us to provide simple and efficient biotin labeling without altering the bioactivity of target proteins. What's more, we have developed multiple biotin labeling approaches to satisfy worldwide customer's requirements.
Creative Biolabs is committed to offering high-quality biotin labeling services for worldwide customers. With advanced technology and proficient scientists, we can assure you both biotinylation efficiency and protein activity preservation. Moreover, our biotin labeling service is reproducible and quality guaranteed. For more detailed information, please feel free to contact us and get a quote. We will get in touch with you within 24 hours and design an optimal method for your project.
Reference
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