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Biochemistry

In vitro Lipid Transfer Assay Authors:  Yan Zhang and Xiaochen Wang, date: 05/05/2013, view: 6264, Q&A: 0
This is a protocol to detect lipid transfer activity of NRF-5, a member of the LPS binding/lipid transfer protein family. The lipid transfer activity is examined by using isotope-labeled cholesterol and liposomes, and tested in two directions (Figure 1): from proteins to liposomes and from liposomes to proteins.
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In vitro Lipid Binding Assay Authors:  Yan Zhang and Xiaochen Wang, date: 05/05/2013, view: 7709, Q&A: 0
This is a protocol to examine in vitro protein-lipid binding using membrane strips coated with various lipids. It has been successfully used to study in vitro interaction between lipids and C. elegans proteins.
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GTP Binding Assays in Arabidopsis thaliana Authors:  Ana Romina Fox and Maria Agustina Mazzella, date: 05/05/2013, view: 4824, Q&A: 0
Signaling through G proteins constitutes an ancient mechanism that functions in the transduction of extracellular signals into intracellular responses. Activation of a proper receptor by a stimuli leads to an exchange of GDP for GTP, the activation of G proteins and the dissociation of Gα-GTP from the Gβγ dimer for the ...
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Cancer Biology

Subcellular Fractionation of Cultured Human Cell Lines Authors:  Zhuoyou Yu, Zhiguang Huang and Maria Li Lung, date: 05/05/2013, view: 26191, Q&A: 3
Subcellular localization is crucial for the proper functioning of a protein. Deregulation of subcellular localization may lead to pathological consequences and result in diseases like cancer. Immuno-fluorescent staining and subcellular fractionation can be used to determine localization of a protein. Here we discuss a protocol to separate the ...
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Developmental Biology

Quantification of Cell Corpses, Cell Death Occurrence, Cell Corpse Duration Authors:  Yan Zhang, Haibin Wang and Xiaochen Wang, date: 05/05/2013, view: 5526, Q&A: 1
During the development of the C. elegans hermaphrodite, 131 of the 1090 somatic cells generated undergo programmed cell death, among which 113 die during embryogenesis starting from 200-cell stage. The apoptotic cells (also called “cell corpses”) appear as highly refractile button-like objects and are easily identified using differential ...
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Immunology

Cell Isolation of Spleen Mononuclear Cells Author:  Benno Weigmann, date: 05/05/2013, view: 14781, Q&A: 0
This method allows you to isolate different subclass mononuclear cells, like B-cells, T cells, CD4+ and CD8+ T, from mouse spleen. By conjugating cells with specific antibodies and subsequently magnetic beads isolation, using the technique from Miltenyi, this allows a high purity.
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Plant Science

Modified Single-Cell Transient Gene Expression Assay in Barley Epidermal Cells Authors:  Shiwei Bai, Cheng Chang, Xinyun Han and Qian-Hua Shen, date: 05/05/2013, view: 5202, Q&A: 0
Transient gene expression via biolistic particle delivery is a widely used technique for gene functional analysis in plants. In this protocol we describe a modified single-cell transient expression assay through transformation with a particle inflow gun of the model PDS-1000/He system (Bio-Rad). This assay was originally optimized for analyzing ...
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Measuring Stomatal Density in Rice Author:  Kensuke Kusumi, date: 05/05/2013, view: 8956, Q&A: 1
The number of stomata on leaves is known to be affected by various environmental factors and intrinsic developmental program. Stomatal density and stomatal index are generally used as indicators of the leaf development and the plant growth. This protocol describes an easy, non-destructive method for preparing imprints of the rice leaf surface ...
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Stem Cell

Single-cell Gene Expression Profiling of Mouse Stem Cells With Fluidigm BiomarkTM Dynamic Array Author:  Ana Sevilla, date: 05/05/2013, view: 10174, Q&A: 3
This protocol describes how to use Fluidigm BiomarkTM 96.96 dynamic arrays for high-throughput expression profiling from single mouse stem cells, assaying up to 96 independent samples with up to 96 quantitative PCR (qPCR) probes (equivalent to 9,216 reactions) in a single experiment. This Dynamic Array contains a network of microfluidic ...
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