Creative Biolabs has established a comprehensive and advanced functional analysis platform for drug discovery. With years of experience in functional analysis, scientists from Creative Biolabs can provide high-quality drug target localization services with numerous strategies.
Target localization is a process to determine where the drug target is located in cells. As the tight correlation of localization and biological function, investigating the localization of targets is crucial for systems biology and targets understanding. The fluorescent protein (FP) fusion, an imaging-based approach, is widely used in target localization as it can provide spatial information on protein location in situ from single cells. Another two radioautographic approaches: thaw-mount and dry-mount are used alone or united with other histochemical approaches for characterizing sites of particular binding and deposition in vivo and in vitro. In addition, gold nanoparticles can be used to position the target molecules as a particulate label. Creative Biolabs can offer unparalleled drug target localization services with diverse approaches to meet all your demands.
Figure 1. Localization of Inn1-GFP in MEN mutants. (Meitinger et al. 2010)
Fluorescent Protein (FP) Fusions for Target Localization
A common imaging-based approach is to express the target product fused to an FP, such as GFP, thereby can temporal study the protein's distribution in its natural state, the living cell. The most available tool to imaging fluorescence is confocal fluorescence microscopy, which can offer actual three-dimensional (3D) optical resolution. In addition, the fusion of the reporter molecule to two or more different tagging locations in each protein, such as the N- and C-termini, is usually required to minimize localization artifacts.
Thaw-mount and Dry-mount Techniques
Thaw-mount and dry-mount techniques are methods permeated in cryo-histology and radioautography.Radioautography with radiolabeled agents is based on the use of traditional tissue fixation and embedding, during which the majority of the labeled compound is translocated and lost. It is the crucial accomplishments that offer a basis for novel applications of target localization. These accomplishments mainly refer to lacking the usual fixation and embedding of several procedures, such as the cutting off of thin frozen sections at low temperatures, conservation of fine structure in freeze-dried and thaw-mounted thin sections, as well as labeled compounds at their initial sites.
Gold Nanoparticles (Au NPs) Technique for Target Localization
Colloidal gold is a collosol suspension of submicrometre-size nanoparticles of gold in a liquid, generally water, and the fluid is commonly a red color (for particles under 100 nm) or blue/purple (for sizeable particles). It can be applied as a particulate label for the localization of target molecules with a variety of modes of electron microscopy, such as scanning and transmission electron microscopy, using both direct and indirect labeling methods.
Creative Biolabs provides various drug discovery services. For more detailed information, please feel free to contact us or directly sent us an inquiry.
References
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