Ribonuclease protection assays

Cytokine profiling has also been measured as a function of changes in cytokine mRNA expression using either reverse transcription polymerase chain reaction (RT-PCR) [87, 91-93] or ribonuclease protection assay (RPA) [94-97], Measurement of cytokine transcripts by RT-PCR revealed that prolonged exposure to TMA induced increased levels of IL-4 mRNA expression compared with treatment with DNCB [87,92-93]. However, expression of the type 1 cytokine IFN-y by DNCB-activated LNC was variable and failed to discriminate between contact and respiratory allergens [87,91,93). A similar profile was observed for freshly isolated tissue analyzed by RPA. This somewhat less... 

Surry DD, McAllister G, Meneses-Lorente G, et al. High-throughput ribonuclease protection assay for the determination of CYP3A mRNA induction in cultured rat hepatocytes. Xenobiotica 1999 29 827-838. 

Choi WS, Machida CA, Ronnekleiv OK. 1995. Distribution of dopamine Dl, D2, and D5 receptor mRNAs in the monkey brain Ribonuclease protection assay analysis. Brain Res Mol Brain Res 31 86-94. 

Richtand NM, Kelsoe JR, Segal DS, Kuczenski R (1995) Regional quantification of Dl, D2, and D3 dopamine receptor mRNA in rat brain using a ribonuclease protection assay. Mol Brain Res 33 97-103. 

Further investigations focused on evaluating the three steps of IFN-y-stimulated class II expression signal transduction and transcription factor activation, CIITA expression, and class II transcription in CMV-infected cells. Ribonuclease protection assays (RPA) and RT-PCR experiments reveal that neither MHC class II nor CIITA RNA is upregulated in response to IFN-y treatment in HCMV-infected ECs at 72h after infection (Miller et al. 1998). Moreover, electrophoretic mobility shift assays (EMSA) demonstrate that STAT-1 homodimers are not... 

Urban levels of air pollution (downtown Sao Paulo) modified the progression of urethane (3 g/ kg)-induced lung tumours in mice (Cury etal. 2000). Urban particles consist of three modes ul-trafine particles, accumulation mode particles (which together form the fine particle mode) and coarse mode particles. Ultrafine particles (those of <100nm diameter) contribute very little to the overall mass, but are very high in number, which in episodic events can reach several hundred thou-sand/cm in the urban air. The hypothesis that ultrafine particles are causally involved in adverse responses seen in sensitive humans is based on several studies summarised by OberdOrster (2001). Timblin etal. (2002) demonstrated the development of dose-related proliferation and apoptosis after exposure of an alveolar epithelial cell line (CIO) to particulate matter or to ultrafine carbon black, a component of particulate matter. Ribonuclease protection assays demonstrated that increases in mRNA levels of the early response protooncogenes c-jun, junB, fra-l, and/ra-2 accompanied cell pro-hferation at low concentrations of particulate matter whereas apoptotic concentrations of particulate... 

Ribonuclease Nj is most stable in neutral or acid media at 37°. It is resistant even to heating at 80° for 2 min in acidic media of pH 2-4, and is fairly unstable in alkaline medium like RNase Ti. It is not as stable as RNase Ti in higher salt concentrations and exhaustive dilution with water makes the enzyme unstable. This enzyme needs a protective protein such as gelatin or bovine serum albumin for assay. 

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