• Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Biochemistry

Protein Translation Study – Label Protein with S35 Methionine in Cells Authors:  Salma Hasan and Isabelle Plo, date: 11/05/2012, view: 19410, Q&A: 2
To follow protein synthesis, cells should be incubated with radioactive amino acid such as [35S] methionine during mRNA translation. Then, the neosynthetized protein will be identified by an autoradiography after immunoprecipitation with a specific antibody and separation on a polyacrylamide denaturing gel.
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Analyze p53 degradation by 35S p53 Pulse Chase Analysis Authors:  Megan Astle, Richard Pearson and Katherine Hannan, date: 11/05/2012, view: 6955, Q&A: 0
p53, is known as the guardian of the genome and as such requires exquisite regulation not only of its abundance but also its activity. The abundance of p53 can be modulated at the level of transcription, translation, and also via its degradation.

This protocol involves 35S metabolic labelling of newly synthesized proteins followed by a ...
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Glucose Production Assay in Primary Mouse Hepatocytes Authors:  Michihiro Matsumoto and Mashito Sakai, date: 11/05/2012, view: 17563, Q&A: 3
Hepatic glucose production is a primary determinant of fasting hyperglycemia in type 2 diabetic patients. Glucagon-cAMP-PKA pathway increases, but insulin-PI3 kinase-Akt pathway suppresses glucose production. This assay aims to evaluate the ability of isolated mouse hepatocytes to release newly synthesized glucose mainly from lactate and pyruvate ...
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In vitro RNA-protein Binding Assay by UV Crosslinking Authors:  Hailong Zhang and Muxiang Zhou, date: 11/05/2012, view: 16903, Q&A: 1
Because covalent bond can form between RNA and its binding proteins after UV irradiation, UV cross-linking is widely used to identify the specific RNA binding proteins. This protocol is described in details as follows.
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Cancer Biology

Establishing Primary Malignant Pleural Mesothelioma (MPM) Cell Cultures Authors:  Mario Cioce, Claudia Canino, Sabrina Strano and Giovanni Blandino, date: 11/05/2012, view: 10428, Q&A: 0
This is a general protocol for the isolation and maintenance of primary MPM cultures as a tool for the identification of Tumor Initiating Cells and early progenitor-targeting drugs (Cioce et al., 2010). Primary cultures can be propagated efficiently for 8-12 weeks and xenotransplanted in NOD/SCID mice while retaining the histofeatures of ...
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Molecular Biology

KMnO4 Footprinting Authors:  Ümit Pul, Reinhild Wurm and Rolf Wagner, date: 11/05/2012, view: 17324, Q&A: 1
The KMnO4 footprinting method offers a rapid and easy way to detect and localize single-stranded regions within a duplex DNA molecule, such as it occurs for instance within an actively transcribing RNA polymerase-DNA complex or during R-loop formation in DNA-RNA hybrid structures. The method is based on the selective oxidation of ...
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Polysome Preparation, RNA Isolation and Analysis Authors:  Hailong Zhang and Muxiang Zhou, date: 11/05/2012, view: 20479, Q&A: 2
During mRNA translation, 40S and 60S ribosomal subunits bind to target mRNA forming into an 80S complex (monosome). This ribosome moves along the mRNA during translational elongation to facilitate tRNA reading codon, where translation is activated and many monosomes can bind the same mRNA simutaneously, which forms polysomes. Polysomes can be ...
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Stem Cell

Isolation of Human Blood Progenitor and Stem Cells from Peripheral Blood by Magnetic Bead Authors:  Salma Hasan and Isabelle Plo, date: 11/05/2012, view: 10671, Q&A: 0
The antigen CD34 is a well-known marker present on human progenitor and stem cells. This protocol explains the isolation of CD34+ cells from peripheral blood using magnetic bead separation technique. The approximate abundance of CD34+ cells in blood is 0.1% of mononuclear cells.
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