Identification and characterization of proteins are ultimately the goal in proteomic analysis. In order to identify a protein trypsin is commonly used to digest protein into peptides which can be analyzed by matrix-assisted laser desorption ionization mass spectrometry (MALDI- ...
In this protocol, we describe an approach in which two-dimensional electrophoresis (2DE)-separated proteins are guanidinated in-gel prior to enzymatic cleavage. In contrast to previously described techniques, this procedure allows the extracted tryptic peptides to be N-term ...
Two-dimensional electrophoresis (2DE) is an excellent technology for the analysis of complex protein mixtures, but it has drawbacks, such as hardly detecting very hydrophobic proteins. Shotgun protein analysis is one of the major technologies used to compensate for the weaknesses of ...
The Drosophila embryonic ventral epidermis has served as a unique tissue for the genetic analysis of patterning. Two types of epidermal cells are easily distinguished: those that secrete short, thick hair-like structures called denticles and cells that only secrete smooth cuticle. De ...
The site-specific recombinase FLP is used in Drosophila to precisely manipulate the genome, in particular, to eliminate gene function by mitotic recombination and to activate transgenes in discrete populations of cells. These approaches are already part of the standard tool kit for stu ...
With the live imaging of embryos, the dynamics of developmental processes, such as tissue remodeling, cell morphogenesis, and, campus protein dynamics can be observed and quantified. This has greatly improved the mechanistic understanding of biological processes. Here we describe ...
The molecular mechanisms controlling Drosophila embryogenesis are among the best-studied examples in animal development. Whereas the formation of developmental pattern in embryos was intensely examined in the past three decades, the cell biological basis of morphogenesis is ...
Drosophila melanogaster imaginal discs are a widely used model system to study signal transduction, developmental, and cell biological processes. An important tool to study these processes is immunolabeling. Fluorochrome-conjugated antibody staining allows the researc ...
The formation of new synapses within neuronal circuits is considered a primary mechanism of long-term synaptic plasticity to allow an increase in synaptic strength. Thus, understanding mechanisms of synapse formation in detail is pivotal for understanding circuit development, as ...
Drosophila pupal (P) wing development entails a series of dynamic developmental events, such as epithelial and glial morphogenesis, that are of outstanding interest to cell biologists. Here, we first describe how to prepare P and prepupal (PP) wings for immunofluorescence microscopy. ...
The regular appearance and the repetitive nature of the Drosophila eye, consisting of several hundred identical multicellular units, the ommatidia, has long served as an invaluable experimental system to study cell-cell interactions, inductive signaling events, cell prolifer ...
Fluorescent in situ hybridization is the standard method for visualizing the spatial distribution of RNA. Although traditional histochemical RNA detection methods suffered from limitations in resolution or sensitivity, the recent development of peroxidase-mediated ty ...
To understand Drosophila development and other genetically controlled processes, it is often desirable to identify differences in gene expression levels. An experimental approach to investigate these processes is to catalog the transcriptome by hybridization of mRNA to DNA mic ...
The fruit fly Drosophila has played a central role in the development of biology during the 20th century. First chosen as a convenient organism to test evolutionary theories soon became the central element in an elaborate, fruitful, and insightful research program dealing with the nature and ...
MicroRNAs (miRNAs), small noncoding RNAs that post-transcriptionally regulate gene expression, are one of the most abundant classes of gene regulators. Yet, little is known about the roles that specific miRNAs play in the development of multicellular organisms. Drosophila provid ...
For many decades, Drosophila has been used as a model system primarily for studies in the fields of genetics and developmental biology. Relatively little attention has been given to the potential of Drosophila as a model system for biochemistry. However, Drosophila embryos as a source for bioc ...
The purification of native protein complexes requires the availability of sufficient amounts of starting material. Drosophila melanogaster embryos have proven to be a rich source for nuclear protein complexes. Here we describe establishment and maintenance of a fly facility for the ...
Drosophila melanogaster is one of the best characterized model systems for genetic analysis. Protein biochemical methods have lagged behind for quite some time but meanwhile have reached a state where protein interaction networks can be elucidated at a similar speed and accuracy as gen ...
Flow cytometry is a powerful technique that allows the researcher to measure fluorescence emissions on a per-cell basis, at multiple wavelengths, in populations of thousands of cells. In this chapter, we outline the use of flow cytometry for the analysis of cells from Drosophila’s imaginal di ...
Given the power of Drosophila genetics, it may seem surprising to discover that many fly researchers are turning to Drosophila cell culture as an experimental system. However, as we will show in this chapter, there are many benefits to be gained by using cell lines as a complement to studies in a tissue and d ...
关于丁香通
公司信息
个人用户
企业机构
