Labeling in situ

Not detected High levels Dual-label in situ hybridization Hrabovszky et al. 2000... 

Luo L-G, Jackson IMD. 1999. Advantage of double labeled in situ hybridization for detecting the effects of glucocorticoids on the mRNAs of protooncogenes and neural peptides TRH in cultured hypothalamic neurons. Brain Res Prot 4 201-208. 

Figure 6.18 ImmunoRCA Assay. Top left A reporter Ab conjugated to an oligonucleotide binds to a test analyte captured on a solid surface by a covalent attachment or by a capture Ab. Top right DNA circle hybridizes to a complementary sequence in the oligonucleotide. Bottom left Resulting complex is washed to remove excess reagents, and the DNA tag is amplified by RCA. Bottom right Amplified product is labeled in situ by hybridization with fluor-labeled oligonucleotides. (From Schweitzer, B. et al, Proc. Natl. Acad. Sci. USA, 97, 10113-10119, 2000. With permission.)...

Figure 15 Representative RP-HPLC Separation of the Tryptic/Chymotryptic Peptides of Bovine Thyrotropin p-Subunit bTSHfS Labeled In Situ with the Fluorescent Reagent 5-[(Iodoacetamidoethyl)amino]naphthalene-l-sulfonic acid (5-l-AEDANS) MI a b...

Bonaventure P, Langlois X, Leysen JE. Co-localization of 5-HT1B- and 5-HT1D receptor mRNA in serotonergic cell bodies in guinea pig dorsal raphe nucleus a double labeling in situ hybridization histochemistry study. Neurosci Lett 1998 254 113-116. 

Relaxin acylated first, then labeled in situ by CT method iodination damage <15%... 

Fig. 1 Human bitter taste receptor cells express subsets of TAS2R genes. Dual labelling in situ hybridization of one taste bud in human circumvallate papillae reveals that cells a and b express hTAS2Rl (upper panel). Of the two cells, cell a contains also hTAS2R10 messenger RNA (mRNA), whereas cell b tests negative for hTAS2R10 mRNA...

Essig-Marcello, J.S. and R.G. Van Buskirk. A double-label in situ cytotoxicity assay using the fluorescent probes neutral red and BCECF-AM. In Vitro Toxicol. 3 219-227, 1990. 

Hohmann AG, Herkenham M (1999b) Localization of central cannabinoid CBI receptor messenger RNA in neuronal subpopulations of rat dorsal root ganglia a double-label in situ hybridization study. Neuroscience 90 923-931... 

Methods for Double- and Single-Label In Situ Hybridization... 

Hohmann, A. G. and Herkenham, M. (2000) Localization of cannabinoid CBl receptor mRNA in neuronal subpopulations of rat striatum a double-label in situ hybridization study. Synapse 37, 71-80. 

Until recently, the application of direct fluorescent labeling in situ hybridization methods was limited, perhaps because the low sensitivity was overemphasized and commercial labeling systems were not available (complex chemistry was required to label the probe or nucleotides). The combination of enzymic labeling techniques and commercial fluorochromized nucleotides, with a better understanding... 

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