Electrophoresis of RNA

Farrell, Robert E.

doi:10.1016/b978-0-12-374727-3.00009-7

Cited by 7 publications

(3 citation statements)

References 45 publications

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“…Briefly, total RNA was extracted using TRIzol (Invitrogen) and cleaned up with DNase I (Sigma Chemical Co.) and subjected to re-extraction when it was necessary. The yield and purity of the extracted RNA were checked using standard protocols of absorbance ratio between 260 and 280 nm, and 1% agarose gel electrophoresis (Farrell 1998a). Furthermore, RIN score of individual samples was determined using Agilent 2100 Bioanalyzer, RNA 6000 Nano LabChip kit, and Agilent 2100 Expert Software (Agilent Technologies, Inc., Santa Clara, CA, USA) (Schroeder et al 2006), and all samples had RIN score O8.0.…”

Section: Rna Extractionmentioning

confidence: 99%

IGF2, IGF binding protein 1, and matrix metalloproteinases 2 and 9 in implantation-stage endometrium following immunoneutralization of vascular endothelial growth factor in the rhesus monkey

Ghosh¹,

Najwa²,

Khan³

et al. 2011

Reproduction

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Blastocyst implantation in the rhesus monkey is inhibited by administration of antibody against vascular endothelial growth factor (VEGF) A during peri-implantation period with no change in the circulatory concentrations of estradiol, progesterone, and VEGF. In this study, we have investigated the effect of administration of a MAB to VEGFA on days 5 and 10 after ovulation upon the mRNA expression, immunopositive protein expression, and immunohistological localization of IGF2, IGF binding protein 1 (IGFBP1) and matrix metalloproteinases (MMPs) 2 and 9 in the implantation-stage endometrium collected on day 13 after ovulation from fecund cycles of rhesus monkeys. The comparison between isotype-matched IgG (control; nZ8)-and VEGF antibody (VEGF Mab; nZ8)-treated animals revealed higher (P!0.05) IGF2 in lacunar and villous syncytiotrophoblasts, trophoblast cell columns, migrating extravillous trophoblast cells, and endovascular trophoblast cells in control animals, but with no change in the various cell types of maternal endometrium between the two groups. No change in IGFBP1 expression in the endometrium was observed between the two groups. MMPs 2 and 9 were detected in syncytiotrophoblast in lacunae and villi, trophoblast cell columns, and extravillous trophoblast cells in control samples. MMP9 transcript expression in maternal endometrium and its immunopositivity in endometrial stroma and trophoblast cells were lower (P!0.05) with no change in MMP2 level in VEGF Mab-exposed samples compared with those in control samples. A functional network involving VEGF, IGF2, and MMP9 in early placental trophoblast cells and maternal endometrium appears to be important for normal placentation.

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Section: Rna Extractionmentioning

confidence: 99%

IGF2, IGF binding protein 1, and matrix metalloproteinases 2 and 9 in implantation-stage endometrium following immunoneutralization of vascular endothelial growth factor in the rhesus monkey

Ghosh¹,

Najwa²,

Khan³

et al. 2011

Reproduction

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“…[2][3][4][5] Many techniques for cellular analysis such as northern blotting, western blotting, fluorescence in situ hybridization (FISH), RT-PCR, DNA microarrays, and electron micro scopy rely on the fixation or lysis of cells. [5][6][7][8][9][10][11] Moreover, as the amount of material from one cell is often insufficient for accurate analysis, a bulk population of cells is used for methods that analyze cell lysates. Information about dynamics of various molecules inside cells and cell-to-cell heterogeneity is often lost in such cases due to ensemble averaging.…”

Section: Introductionmentioning

confidence: 99%

Nucleic‐Acid Structures as Intracellular Probes for Live Cells

2019

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The chemical composition of cells at the molecular level determines their growth, differentiation, structure, and function. Probing this composition is powerful because it provides invaluable insight into chemical processes inside cells and in certain cases allows disease diagnosis based on molecular profiles. However, many techniques analyze fixed cells or lysates of bulk populations, in which information about dynamics and cellular heterogeneity is lost. Recently, nucleic‐acid‐based probes have emerged as a promising platform for the detection of a wide variety of intracellular analytes in live cells with single‐cell resolution. Recent advances in this field are described and common strategies for probe design, types of targets that can be identified, current limitations, and future directions are discussed.

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“…Due to the potential for artifacts introduced by contaminating RNases, care should be taken to follow best practices, such as the use of RNase-free solutions and reagents and/or using DEPC-treatment where appropriate (Farrell, 2010). Standard materials and reagents for urea-polyacrylamide gel electrophoresis are required; we use the National Diagnostics system but such gels can be prepared using standard methods (Sambrook and Russell, 2006).…”

Section: Introductionmentioning

confidence: 99%

RNA Degradation Assay Using RNA Exosome Complexes, Affinity-purified from HEK-293 Cells

Domanski

LaCava

2017

BIO-PROTOCOL

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The RNA exosome complex plays a central role in RNA processing and regulated turnover. Present both in cytoplasm and nucleus, the exosome functions through associations with ribonucleases and various adapter proteins (reviewed in [Kilchert et al., 2016]). The RNA exosome-associated EXOSC10 protein is a distributive, 3’–5’ exoribonuclease. The following protocol describes an approach to monitor the ribonucleolytic activity of affinity-purified EXOSC10-containing RNA exosomes, originating from HEK-293 cells, as reported in (Domanski et al., 2016) and further detailed in the companion bio-protocol to this one (Domanski and LaCava, 2017).

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Electrophoresis of RNA

Cited by 7 publications

References 45 publications

IGF2, IGF binding protein 1, and matrix metalloproteinases 2 and 9 in implantation-stage endometrium following immunoneutralization of vascular endothelial growth factor in the rhesus monkey

IGF2, IGF binding protein 1, and matrix metalloproteinases 2 and 9 in implantation-stage endometrium following immunoneutralization of vascular endothelial growth factor in the rhesus monkey

Nucleic‐Acid Structures as Intracellular Probes for Live Cells

RNA Degradation Assay Using RNA Exosome Complexes, Affinity-purified from HEK-293 Cells

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