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Binding Stoichiometry of an Antibody Fragment
Binding Stoichiometry of an Antibody Fragment
3 Feb 2025Antibody fragments, especially Fab-fragments, have emerged as a tool to facilitate the crystal growth and improve crystal quality of membrane proteins. This application note demonstrates the measurement of binding stoichiometry using the powerful method of multi-angle light scattering in combination with size exclusion chromatography (MALS-SEC). This information is instrumental in further biophysical studies and co-crystallization experiments. Additional information such as weight fraction of protein and detergent in the protein-detergent micelle are obtained in the same experiment when using both UV and RI-detectors.
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Size Exclusion ChromatographySize exclusion chromatography (SEC) is a liquid chromatographic technique used to separate and identify molecules based on their size, including polymers, proteins, nanoparticles & nucleic acid. SEC is further subdivided into gel filtration chromatography (GFC) and gel permeation chromatography (GPC) when an aqueous or organic mobile phase is used respectively. HPLC-based SEC systems and columns are available for analytical separation alongside SEC spin columns or filters to prep samples for many protocols. Find the best size exclusion chromatography equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.AntibodiesAntibodies are used in techniques such as confocal and fluorescence microscopy, flow cytometry, ELISA, ELISPOT, immunohistochemistry, western blotting and immunopreciptation. Select specific antigen reactivity, high specific affinity, low non-specific binding, monoclonal or polyclonal, primary or secondary antibodies and associated conjugates such as an enzyme or dye for visualization.Protein CrystallographyProtein crystallization is the process of crystallizing purified proteins for 3D structure analysis by x-ray crystallography. The main methods of protein crystallization include sitting drop, hanging drop and microbatch. It is important to control parameters such as pH, temperature and concentration. Following crystallization, detectors and software are used for data collection and analysis.DNA Fragment SizingMolecular Weight AnalysisMolecular weight analysis determines the size and structure of biomolecules, essential in protein characterization, polymer studies, and drug development. Techniques like mass spectrometry ensure precise results. Compare solutions for molecular weight analysis in our peer-reviewed product directory.Antibody ArraysSECSize exclusion chromatography (SEC) separates molecules based on size, ideal for protein purification, polymer analysis, and biopharma applications. SEC ensures precise characterization of biomolecules for research and industry. Explore peer-reviewed SEC products and compare options for your lab.Crystallization