Protocols in Current Issue
    mRNA Stability Assay Using Transcription Inhibition by Actinomycin D in Mouse Pluripotent Stem Cells
    Authors:  Madara Ratnadiwakara and Minna-Liisa Änkö, date: 11/05/2018, view: 5135, Q&A: 0
    [Abstract] Gene expression is regulated through multiple steps at both transcriptional and post-transcriptional levels. The net abundance of mature mRNA species in cells is determined by the balance between transcription and degradation. Thus, the regulation of mRNA stability is a key post-transcriptional event that can greatly affect the net level of mRNAs ...
    RNA Immunoprecipitation Assay to Determine the Specificity of SRSF3 Binding to Nanog mRNA
    Authors:  Madara Ratnadiwakara and Minna-Liisa Änko, date: 11/05/2018, view: 1571, Q&A: 0
    [Abstract] Mammalian cells express hundreds of RNA binding proteins (RBPs) that are essential regulators of RNA metabolism. RBP activity plays a central role in the control of gene expression programs and identification of RNA-protein interactions is critical for comprehensive understanding of gene regulation in cells. In recent years, various tools and ...
    Isolation of LYVE-1+ Endothelial Cells from Mouse Embryos
    Author:  Patrick Crosswhite, date: 08/05/2018, view: 1537, Q&A: 0
    [Abstract] Lymphatic vessel endothelial hyaluronan receptor 1, or LYVE-1, is a type 1 integral membrane glycoprotein expressed by lymphatic endothelial cells (LECs). LYVE-1 is commonly used as a biological marker to visually distinguish developing lymphatic vessels from blood endothelial cells (arteries or veins). As our understanding of lymphatic biology is ...
    FACS-based Glucose Uptake Assay of Mouse Embryonic Fibroblasts and Breast Cancer Cells Using 2-NBDG Probe
    Authors:  Shengli Dong and Suresh K Alahari, date: 04/20/2018, view: 3537, Q&A: 0
    [Abstract] This is a flow cytometry-based protocol to measure glucose uptake of mouse embryonic fibroblasts (MEFs) and breast cancer cells in vitro. The method is a slightly modified and updated version as previously described (Dong et al., 2017). Briefly, the target cells are incubated with the fluorescently tagged ...
    Histochemical Staining of Collagen and Identification of Its Subtypes by Picrosirius Red Dye in Mouse Reproductive Tissues
    [Abstract] Collagen is one of the foremost components of tissue extracellular matrix (ECM). It provides strength, elasticity and architecture to the tissue enabling it to bear the wear and tear from external factors like physical stress as well as internal stress factors like inflammation or other pathological conditions. During normal pregnancy or pregnancy ...
    Isolation and Culturing of Rat Primary Embryonic Basal Forebrain Cholinergic Neurons (BFCNs)
    Authors:  Wei Xu and Chengbiao Wu, date: 07/20/2017, view: 3190, Q&A: 0
    [Abstract] The basal forebrain is located close to the medial and ventral surfaces of the cerebral hemispheres that develop from the sub-pallium. It regulates multiple processes including attention, learning, memory and sleep. Dysfunction and degeneration of basal forebrain cholinergic neurons (BFCNs) are believed to be involved in many disorders of the ...
    Reprogram Murine Epiblast Stem Cells by Epigenetic Inhibitors
    Authors:  Hui Zhang and Yali Dou, date: 03/05/2017, view: 3639, Q&A: 0
    [Abstract] Pluripotent stem cells in the naïve state are highly useful in regenerative medicine and tissue engineering. A robust reprogramming of the primed murine Epiblast Stem Cells (EpiSCs) to naïve pluripotency is feasible via chemical-only approach. This protocol described a method to reprogram murine EpiSCs by MM-401 treatment, which blocks histone ...
    Mouse Oocyte Isolation, Cultivation and RNA Microinjection
    Authors:  Anna Tetkova and Marketa Hancova, date: 02/05/2016, view: 9123, Q&A: 0
    [Abstract] Mammalian oocyte is a highly specialized cell, characterized by synthesis and storage of maternal proteins and RNAs that contributes to the meiotic cell cycle and early embryo development. The fully grown oocyte is transcriptionally quiescent and utilizes only transcripts synthesized and stored during the growing phase. Mouse oocytes are often ...
    Single Molecule RNA FISH in the Mammalian Oocyte
    Author:  Denisa Jansova , date: 12/05/2015, view: 5737, Q&A: 0
    [Abstract] RNA fluorescence in situ hybridization is a method to localize and measure gene expression in individual cell or tissue. Using multiple specific fluorescently labeled oligonucleotides greatly increases signal-to-noise ratio and thus enables detection of single RNA molecule. Around forty different DNA oligonucleotides designed to common ...
    Hypertrophy Analysis and Quantification in Embryonic Cardiomyocites
    Authors:  Guillermo Luxán and José Luis de la Pompa, date: 10/20/2013, view: 6218, Q&A: 0
    [Abstract] Myocardial growth goes from proliferation to hypertrophy during development. The measurement of the relative cell area provides information of cardiomyocyte hypertrophy, which is ideal for studying myocardial development.

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