In-Situ Hybridization to Sections (Nonradioactive)
In-situ hybridization (ISH) takes advantage of the ability of mRNA within a cell to hybridize with exogenously applied complementary RNA (riboprobes) or DNA molecules. This interaction is visualized by labeling the applied nucleic acid probe with a detectable molecule (radioactive, such as 35S, or nonradi-oactive, such as digoxygenin [DIG]). The technique allows patterns of gene expression to be visualized in many tissues or cell types simultaneously. Nonradioactive ISH has several advantages over radioactive ISH. It lacks the biohazards associated with the use of radioisotopes, it takes days rather than weeks to get a visible signal, and it is cheaper. Using nonradioactive detection, the degree of cellular resolution is significantly improved over that achieved with autoradiography. Another advantage of nonradioactive ISH is that it can be performed in combination with other assays with relative ease, to compare mRNA and protein distribution in the same tissues (1),...
KeywordsColor Detection Sheep Serum Resuspend Pellet Blue Tetrazolium Chloride Alkaline Phosphatase Streptavidin
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