细胞功能测定

The successful treatment of primary and secondary bone tumors in a huge number of cases remains one of the major unsolved challenges in modern medicine. Malignant primary bone tumor growth predominantly occurs in younger people, whereas older people predominantly suffer from seconda ...

Polyamines are aliphatic polycations that function in key cellular processes such as growth, differentiation, and macromolecular biosynthesis. Intracellular polyamines pools are maintained from de novo synthesis and from transport of polyamines from the extracellular m ...

Acrolein is an α,β-unsaturated aldehyde with enormous capacity of reaction, occurs in the air like a pollutant, but it is (we know now) an important lipid peroxidation product as well. The compound is one of the several aldehydes produced from fatty acid oxidation, although it is particularly imp ...

Methods are presented for the production, affinity purification and analysis of plasmid DNA (pDNA). Batch fermentation is used for the production of the pDNA, and expanded bed chromatography, via the use of a dual affinity glutathione S-transferase (GST) fusion protein, is used for the captu ...

The zona pellucida is an extracellular glycoprotein coat that surrounds the oocyte and, in many mammals, provides the first barrier sperm must penetrate for fertilization to occur. The molecular interactions between the sperm and zona pellucida appear to be relatively species-speci ...

Many ion channels, particularly potassium channels, are regulated by intracellular substances, such as nucleotides or Ca2+. These modulators are washed out of the cell during standard whole-cell patch clamp recordings, or maintained at a particular concentration if they are includ ...

This section focuses on the application of differential display in gaining insight into cDNAs upregulated in association with a disease state by using direct in vivo samples. Although the majority of differential display studies have focused on in vitro models, a growing number of laborat ...

The technology for immobilization of cells and enzymes evolved steadily for the first 25 years of its existence (1), but in recent years it has reached a plateau, if not a slight decline. However, the expansion of biotechnology, and the expected developments that will accrue from advances in genetic ...

In blood vessel development (vasculogenesis) smooth muscle cells (SMCs) are derived by differentiation of mesenchymal cells under the influence of mediators secreted by the endothelial cells (ECs) composing newly formed vessels. In angiogenesis, SMCs can be formed in the same way or by p ...

Expression of recombinant matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in mammalian cells is an important step in their functional characterization. Also, transient transfection analysis of promoter constructs driving CAT or luc ...

Immobilized metal affinity chromatography (IMAC) is a common place technique in modern protein purification. IMAC is distinct from most other affinity chromatography technologies in that it can operate on a native, unmodified protein without the need for a specialized affinity “tag” ...

Metal affinity precipitation has been successfully developed as a simple purification process for the proteins that have affinity for the metal ions. The copolymers of vinylimidazole with N-isopropylacrylamide are easily synthesized by radical polymerization. When loaded w ...

With the near completion of the human genome sequencing effort, it is now possible to analyze the expression of the entire human gene complement. However, a major obstacle in performing such analysis is the ability to successfully generate enough cDNA or amplified RNA from a limited number of cel ...

Quantification of the expression of individual genes can reveal much concerning the processes occurring within a cell. In the vast majority of cases, activation or repression of a gene is indicative of altered utilization of the pathway or process in which it functions. Although microarray ...

Comparative Genomic Hybridization (CGH) using metaphase chromosome spreads to screen all human chromosomes for aneuploidy in preimplantation embryos is hindered by the time required to perform the analysis. DNA microarrays manufactured to date are not able to analyze the very limi ...

Laser capture microdissection (LCM) can be used to obtain single cells or a homogeneous population of cells for molecular analysis. This approach becomes even more powerful when it is combined with immunocytochemical staining using specific antibodies to label the cells of interest be ...

Microarrays containing oligonucleotide mutation probes are emerging as useful platforms for the diagnosis of genetic disease. Herein, we describe the development and validation of an inhouse microarray suitable for the diagnosis of common cystic fibrosis (CF) mutations in low-t ...

Because metabolism is fundamental to the successful functioning of a cell, perturbations in metabolism can signal compromised viability. For the assessment of viability, analyses not only must be sensitive enough to be employed at the single cell level, but also noninvasive. Ultramic ...

Fluorescence in situ hybridization (FISH) is the technique of choice for preimplantation genetic diagnosis (PGD) selection of female embryos in families with X-linked disease, for which there is no mutation-specific test. FISH with target-specific DNA probes is also the primary tech ...

We provide an overview of the methodology involved in single cell polymerase chain reaction (PCR), especially for single lymphocytes or cultured lymphoblasts and blastomeres. The first step toward single cell PCR is isolation of single cells; the protocols given can be carried out using ba ...