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Methods and Protocols, Volume 3, Issue 2

June 2020 - 21 articles

Cover Story: The analysis of glycosylphosphatidylinositol (GPI)-anchored receptor distribution and dynamics in live cells is challenging because their clusters exhibit subdiffraction-limited sizes and are highly dynamic. We compared three approaches to receptor labeling and found that the enzyme-mediated site-specific fluorescence labeling of receptor modified with a short peptide tag has several advantages over labeling by fluorescent proteins or antibodies. We successfully detected the Förster resonance energy transfer (FRET) of GPI-anchored receptors, T-cadherin, and ephrin-A1 without ligands and showed in real time that adiponectin induces stable T-cadherin cluster formation. Here, we present the practical aspects of labeling and the heteroFRET measurements of GPI-anchored receptors to study their dynamics on a plasma membrane in live cells. View this paper.
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Articles (21)

  • Article
  • Open Access
18 Citations
5,204 Views
11 Pages

Validation of a Simple HPLC–UV Method for the Determination of Monomers Released from Dental Resin Composites in Artificial Saliva

  • Elisavet-Ioanna Diamantopoulou,
  • Orfeas-Evanggelos Plastiras,
  • Petros Mourouzis and
  • Victoria Samanidou

Bisphenol-A (BPA), bisphenol A glycerolate dimethacrylate (Bis-GMA), triethylene glycol dimethacrylate (TEGDMA), and urethane dimethacrylate (UDMA) are organic monomers that can be released from dental composites into the oral cavity. Over specific c...

  • Article
  • Open Access
18 Citations
6,610 Views
15 Pages

OCTAVVS: A Graphical Toolbox for High-Throughput Preprocessing and Analysis of Vibrational Spectroscopy Imaging Data

  • Carl Troein,
  • Syahril Siregar,
  • Michiel Op De Beeck,
  • Carsten Peterson,
  • Anders Tunlid and
  • Per Persson

Modern vibrational spectroscopy techniques enable the rapid collection of thousands of spectra in a single hyperspectral image, allowing researchers to study spatially heterogeneous samples at micrometer resolution. A number of algorithms have been d...

  • Article
  • Open Access
5 Citations
5,315 Views
14 Pages

The Making of Transgenic Drosophila guttifera

  • Mujeeb Shittu,
  • Tessa Steenwinkel,
  • Shigeyuki Koshikawa and
  • Thomas Werner

The complex color patterns on the wings and body of Drosophila guttifera (D. guttifera) are emerging as model systems for studying evolutionary and developmental processes. Studies regarding these processes depend on overexpression and downregulation...

  • Protocol
  • Open Access
3 Citations
4,167 Views
9 Pages

In this protocol, we introduced a method of measuring mitochondrial dysfunction to confirm the epithelial–mesenchymal transition (EMT) in pancreatic cancer cells under a hypoxic environment. There are many expertized and complicated methods to...

  • Article
  • Open Access
4 Citations
4,832 Views
21 Pages

Analysis of GPI-Anchored Receptor Distribution and Dynamics in Live Cells by Tag-Mediated Enzymatic Labeling and FRET

  • Maria N. Balatskaya,
  • Alexandra I. Baglay,
  • Yury P. Rubtsov and
  • George V. Sharonov

The analysis of glycosylphosphatidylinositol (GPI)-anchored receptor distribution and dynamics in live cells is challenging, because their clusters exhibit subdiffraction-limited sizes and are highly dynamic. However, the cellular response depends on...

  • Protocol
  • Open Access
26 Citations
7,574 Views
9 Pages

Novel bone substitute materials need to be evaluated in terms of their osteogenic differentiation capacity and possible unwanted cytotoxic effects in order to identify promising candidates for the therapy of bone defects. The activity of alkaline pho...

  • Article
  • Open Access
5 Citations
3,971 Views
15 Pages

Quantifying Physical Thrombus Characteristics on Cardiovascular Biomaterials Using MicroCT

  • Avi Gupta,
  • Claire M. Johnston,
  • Monica T. Hinds and
  • Deirdre E. J. Anderson

Hemocompatibility is a critical consideration when designing cardiovascular devices. Methods of assessing hemocompatibility range from in vitro protein adsorption and static platelet attachment to in vivo implantation. A standard preclinical assessme...

  • Technical Note
  • Open Access
28 Citations
3,758 Views
16 Pages

Quantitative trait loci (QTL) are genomic regions associated with phenotype variation of quantitative traits. To date, a total of 313 QTL for 31 quantitative traits have been reported in 14 studies on flax. Of these, 200 QTL from 12 studies were iden...

  • Protocol
  • Open Access
56 Citations
11,950 Views
12 Pages

An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length

  • Mugdha V. Joglekar,
  • Sarang N. Satoor,
  • Wilson K.M. Wong,
  • Feifei Cheng,
  • Ronald C.W. Ma and
  • Anandwardhan A. Hardikar

Telomeres represent the nucleotide repeat sequences at the ends of chromosomes and are essential for chromosome stability. They can shorten at each round of DNA replication mainly because of incomplete DNA synthesis of the lagging strand. Reduced rel...

  • Protocol
  • Open Access
3 Citations
4,586 Views
11 Pages

Exploring Patient Views and Acceptance of Multiparametric Magnetic Resonance Imaging for the Investigation of Suspected Prostate Cancer (the PACT Study): A Mixed-Methods Study Protocol

  • Joseph M. Norris,
  • Veeru Kasivisvanathan,
  • Clare Allen,
  • Rhys Ball,
  • Alex Freeman,
  • Maneesh Ghei,
  • Alex Kirkham,
  • Hayley C. Whitaker,
  • Daniel Kelly and
  • Mark Emberton

BACKGROUND: The introduction of multiparametric magnetic resonance imaging (mpMRI) has improved the diagnosis of suspected prostate cancer, accurately risk-stratifying men before a biopsy. However, pre-biopsy mpMRI represents a significant deviation...

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Methods Protoc. - ISSN 2409-9279