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- W936624145 abstract "Publisher Summary This chapter describes the role of nuclease probing in determining RNA–protein and RNA–ribonucleoprotein interactions. A radiolabeled precursor is incubated with an extract or fraction that contains the ribonucleoprotein (RNP) or protein being tested for binding. An active extract should be used under conditions that allow the processing reaction of interest to occur. The specificity and titer of the antibody used for immunoprecipitation are very important. For most protection–immunoprecipitation experiments, either mouse monoclonal antibodies or human sera derived from patients with autoimmune diseases have been used. Deviation from an established protocol may require retitration of the amount of nuclease needed to achieve adequate digestion of the precursor. An excellent way to test whether a particular RNP is involved in the protection–immunoprecipitation reaction is to target its RNA for cleavage, using a complementary oligodeoxynucleotide and RNase H, which cleaves the RNA component of a RNA–DNA hybrid. Difficulties in interpretation arise if a fragment can be immunoprecipitated to a lesser extent with control antibodies." @default.
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- W936624145 date "1989-01-01" @default.
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- W936624145 title "[31] Determination of RNA-protein and RNA-ribonucleoprotein interactions by nuclease probing" @default.
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- W936624145 doi "https://doi.org/10.1016/0076-6879(89)80117-x" @default.
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