alexa Analysis of cell cycle by flow cytometry.
Chemical Engineering

Chemical Engineering

Journal of Bioprocessing & Biotechniques

Author(s): Pozarowski P, Darzynkiewicz Z, Pozarowski P, Darzynkiewicz Z

Abstract Share this page

Abstract Described are four widely used procedures to analyze the cell cycle by flow cytometry. The first two are based on univariate analysis of cellular DNA content following cell staining with either propidium iodide (PI) or 4',6'-diamidino-2-phenylindole (DAPI) and deconvolution of the cellular DNA content frequency histograms. This approach reveals distribution of cells in three major phases of the cycle (G1 vs S vs G2/M) and makes it possible to detect apoptotic cells with fractional DNA content. The third approach is based on the bivariate analysis of DNA content and proliferation-associated proteins. The expression of cyclin D, cyclin E, cyclin A, or cyclin B1 vs DNA content is presented as an example. This approach allows one to distinguish, for example, G0 from G1 cells, identify mitotic cells, or relate expression of other intracellular proteins to the cell cycle position. The fourth procedure relies on the detection of 5'-bromo-2'-deoxyuridine (BrdU) incorporation to label the DNA-replicating cells. This article was published in Methods Mol Biol and referenced in Journal of Bioprocessing & Biotechniques

Relevant Expert PPTs

Relevant Speaker PPTs

  • Alexandra Emelyanova
    Released-active antibodies are innovative products for the effective management of severe respiratory viral infections
    PPT Version | PDF Version
  • Azeez L
    Precursors influencing tropospheric ozone formation and apportionment in three districts of Ilupeju Industrial Estate, Lagos
    PPT Version | PDF Version
  • Chang-Hung Chou
    Diversified natural products in Rhododendron formosanum reveal allelochemical and pharmaceutical properties
    PPT Version | PDF Version
  • Yi-Cheng Hu
    Detection of a negative correlation between prescription of Chinese herbal products containing coumestrol, genistein or daidzein and risk of subsequent endometrial cancer among tamoxifentreated female breast cancer survivors in Taiwan between 1998 and 2008: A population-based study
    PPT Version | PDF Version
  • Carsten Worsoe
    The value of the “simulated study” as a tool to predict actual leachables in parenteral drug products
    PPT Version | PDF Version
  • Diane Paskiet
    Qualifi cation of extractables & leachables from container closure systems in drug products- Introduction to the Product Quality Institute (PQRI) recommendations
    PPT Version | PDF Version
  • Neervalur V Raghavan
    Systematic approach to development of aqueous drug formulation and drug device combination injectable products and challenges
    PPT Version | PDF Version
  • Jianfeng Hong
    Extractable and leachable studies of parenteral infusion and transfusion products
    PPT Version | PDF Version
  • Heidi Schalchli
    Natural products of Anthracophyllum discolor: Ligninolytic enzymes and antifungal volatile compounds
    PPT Version | PDF Version
  • Hazel Gorham
    Challenges in demonstrating biosimilarity and interchangeability of biosimilar products
    PPT Version | PDF Version
  • T. B. Karegoudar
    Purification and Characterization of Amidase from Paracoccus sp. SKG: Utilization of amidase inhibited whole cells for bioconversion of acrylonitrile to acrylamide
    PPT Version | PDF Version
  • Uyoh Edak Aniedi
    PROCESS OPTIMIZATION OF BIOETHANOL PRODUCTION FROM PEELS OF CASSAVA CULTIVARS USING DIFFERENT MICROBIAL INNOCULANTS
    PPT Version | PDF Version
  • Mikael Bjerg Caspersen
    Innovative albumin based technology for half-life extension and optimization of Biotherapeutics
    PPT Version | PDF Version
  • Monica C Chuong
    Bioprocess Development Upstream and Downstream Technologies
    PPT Version | PDF Version
  • Yosef Yarden
    Classically, the 3’untranslated region (3’UTR) is that region in eukaryotic protein-coding genes from the translation termination codon to the polyA signal. It is transcribed as an integral part of the mRNA encoded by the gene. However, there exists another kind of RNA, which consists of the 3’UTR alone, without all other elements in mRNA such as 5’UTR and coding region. The importance of independent 3’UTR RNA (referred as I3’UTR) was prompted by results of artificially introducing such RNA species into malignant mammalian cells. Since 1991, we found that the middle part of the 3’UTR of the human nuclear factor for interleukin-6 (NF-IL6) or C/EBP gene exerted tumor suppression effect in vivo. Our subsequent studies showed that transfection of C/EBP 3’UTR led to down-regulation of several genes favorable for malignancy and to up-regulation of some genes favorable for phenotypic reversion. Also, it was shown that the sequences near the termini of the C/EBP 3’UTR were important for its tumor suppression activity. Then, the C/EBP 3’UTR was found to directly inhibit the phosphorylation activity of protein kinase CPKC in SMMC-7721, a hepatocarcinoma cell line. Recently, an AU-rich region in the C/EBP 3’UTR was found also to be responsible for its tumor suppression. Recently we have also found evidence that the independent C/EBP 3’UTR RNA is actually exists in human tissues, such as fetal liver and heart, pregnant uterus, senescent fibroblasts etc. Through 1990’s to 2000’s, world scientists found several 3’UTR RNAs that functioned as artificial independent RNAs in cancer cells and resulted in tumor suppression. Interestingly, majority of genes for these RNAs have promoter-like structures in their 3’UTR regions, although the existence of their transcribed products as independent 3’UTR RNAs is still to be confirmed. Our studies indicate that the independent 3’UTR RNA is a novel non-coding RNA species whose function should be the regulation not of the expression of their original mRNA, but of some essential life activities of the cell as a whole.
    PPT Version | PDF Version

Recommended Conferences

  • 17th Euro Biotechnology Congress
    September 25-27, 2017 Berlin, Germany
  • 2nd World Biotechnology Congress
    December 04-06, 2017 Sao Paulo, Brazil

Relevant Topics

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

 
© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
adwords