Cell type–specific mRNA purification by translating ribosome affinity purification (TRAP)
نویسندگان
چکیده
منابع مشابه
TRAP-rc, Translating Ribosome Affinity Purification from Rare Cell Populations of Drosophila Embryos.
Measuring levels of mRNAs in the process of translation in individual cells provides information on the proteins involved in cellular functions at a given point in time. The protocol dubbed Translating Ribosome Affinity Purification (TRAP) is able to capture this mRNA translation process in a cell-type-specific manner. Based on the affinity purification of polysomes carrying a tagged ribosomal ...
متن کاملDevelopment of translating ribosome affinity purification for zebrafish.
The regulation of transcription and translation by specific cell types is essential to generate the cellular diversity that typifies complex multicellular organisms. Tagging and purification of ribosomal proteins has been shown to be an innovative and effective means of characterizing the ribosome bound transcriptome of highly specific cell populations in vivo. To test the feasibility of using ...
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The glycine receptor of rat spinal cord was solubilized with the nonionic detergent Triton X-100 and subsequently purified by affinity chromatography on aminostrychnine-agarose and wheat germ agglutininSepharose. An overall purification of 1950-fold was achieved. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and mercaptoethano1 revealed three glycine receptor-asso...
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The preparation of a number of agarose and polyacrylamide bead derivatives useful in the purification of proteins by afhnity chromatography is described. These techniques permit (a) the attachment of ligands to the gel through extended hydrocarbon chains which place the ligand at varying distances from the gel matrix backbone; (b) the covalent attachment of ligands to agarose or polyacrylamide ...
متن کاملProtein Purification by Affinity Chromatography
The preparation of a number of agarose and polyacrylamide bead derivatives useful in the purification of proteins by afhnity chromatography is described. These techniques permit (a) the attachment of ligands to the gel through extended hydrocarbon chains which place the ligand at varying distances from the gel matrix backbone; (b) the covalent attachment of ligands to agarose or polyacrylamide ...
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ژورنال
عنوان ژورنال: Nature Protocols
سال: 2014
ISSN: 1754-2189,1750-2799
DOI: 10.1038/nprot.2014.085