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细胞功能测定

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Immobilization of Enzymes on Glyoxyl Agarose: Strategies for Enzyme Stabilization by Multipoint Attachment

Immobilization of enzymes on pre-existing supports containing small aliphatic aldehyde groups (e.g., glyoxyl-agarose) presents a set of interesting features from both a basic and a more practical point of view. The chemistry of this immobilization method is quite different from more p ...

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Activation of Rayon/Polyester Cloth for Protein Immobilization

The immobilization of proteins is an essential step in constructing affinity purification or enzyme reactor systems. For these biotechnological processes, immobilized proteins can be used in either batchwise or column operations. Commonly utilized insoluble supports for pr ...

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Entrapment of Enzymes and Cells in Poly (2-Hydroxyethyl Methacrylate) Supports

Immobilization of enzymes and cells has been widely investigated for many years, and numerous studies and reviews have appeared in the literature. In fact, immobilization offers the possibility to improve stability of the catalyst and is also considered the simplest and most effective m ...

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Entrapment in Calcium Alginate

Entrapment of enzymes and/or cells in alginate is one of the simplest methods of immobilization. Alginates are available commercially as water-soluble sodium alginates, and have been used for more than 65 years in the food and pharmaceutical industries as thickening, emulsifying, film f ...

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Immobilization in Carrageenans

Immobilization in carrageenans is a method of gel entrapment. It is one of the most widely used methods for cell immobilization because it is cheap, simple, and reproducible with mild conditions during the immobilization. The major advantages of the entrapment method are high cell density, m ...

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Visual Assessment of Enzyme Immobilization

Immobilization of enzymes is a basic technique that is well established in very different fields of both research and industry. The actual knowledge of the basic principles for immobilizing enzymes is so broad that one can design the immobilized enzyme to accomplish a given task or to show prope ...

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Enzyme Immobilization on Nylon

Nylons are a family of linear condensation polymers, involving the repeated alkane segments bound by secondary amide linkages. The different types of nylons available commercially are named and classified according to the number of carbon atoms in their monomeric units (i.e., nylon 6,6 is o ...

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Transition Metal Methods for Immobilization of Enzymes and Cells

The use of transition metal chemistry for the immobilization of biological molecules is long established and well documented (1–17). This method is based on the chelation properties of transition metals, namely titanium(IV) and zirconium(IV), which seem particularly attractive on a ...

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Use of Divalent Metal Ions Chelated to Agarose Derivatives for Reversible Immobilization of Proteins

Immobilization of a protein through coordinate bonds formed with divalent metal ions (e.g., Me(II), Cu(II)) is becoming an attractive alternative to covalent coupling chemistries. This is primarily a result of the reversible nature of the immobilization, because the protein may be easily ...

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Molecular Single-Cell PCR Analysis of Rearranged Immunoglobulin Genes As a Tool to Determine the Clonal Composition of Normal and Malignant Human B Ce

Owing to the nearly limitless diversity of immunoglobulin (Ig) variable-region gene rearrangements, such rearrangements represent ideal clonal markers for B-lineage cells. This chapter describes an approach to isolate single cells from frozen tissue sections by microdisse ...

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Using TUNEL in Combination with an Active Caspase-3 Immunoassay to Identify Cells Undergoing Apoptosis in Preimplantation Mammalian Embryos

The term apoptosis was coined in a now classic paper by Kerr et al. in 1972 (1), which summarized the morphological features observed in electron micrographs of cells dying by a process that had been previously termed shrinkage necrosis. It was noted that these features, such as cell shrinkage, nucle ...

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Acquisition, Display, and Analysis of Digital Three-Dimensional Time-Lapse (Four-Dimensional) Data Sets Using Free Software Applications

The ability to capture and visualize information throughout the volume of a living three-dimensional (3-D) specimen as it changes over time (four-dimensional imaging) has been the goal of many biological researchers. In practice, this is usually accomplished by gathering a series of 3-D da ...

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Active Protease Mapping in 2DE Gels

Proteases act as the molecular mediators of many vital biological processes. To understand the function of each protease, it needs to be separated from other proteins and characterized in its natural, biologically active form. In the method described in this chapter, proteases in a biologi ...

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Detection of Human Papillomavirus DNA in Sperm Using Polymerase Chain Reaction

Human papillomavirus (HPV) is a highly prevalent sexually transmitted virus that causes genital warts in men and women and is associated with an increased risk of genital tract neoplasia in women (1,2). Nucleic acid amplification techniques such as the polymerase chain reaction (PCR) have ...

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Spermatogonial Stem Cells

Spermatogenesis is a continuous, highly organized process comprised of sequential steps of cell proliferation and differentiation (1). In male mammals, spermatogenesis proceeds for the reproductive lifetime of the animals. The continuation of this process depends on a pool of spe ...

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Molecular and Cellular Approaches for Establishment of an In Vitro Spermatogenesis System

Spermatogenesis is a complex, well-organized process including proliferation, differentiation, apoptosis, and morphogenesis through which differentiating daughter cells of spermatogonial stem cells develop into mature sperm (1,2). In men, this process requires approx ...

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Methods to Study Transcription-Coupled Repair in Chromatin

Transcription-coupled repair (TCR) is a sub-pathway of nucleotide excision repair that allows for the enhanced repair of the transcribed strand of active genes. A classical method to study DNA repair in vivo consists in the molecular analysis of UV-induced DNA damages at specific loci. Cells ...

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Cytometric Analysis of DNA Damage: Phosphorylation of Histone H2AX as a Marker of DNA Double-Strand Breaks (DSBs)

Phosphorylation of histone H2AX on Ser 139 is a sensitive reporter of DNA damage, particularly if the damage involves induction of DNA double-strand breaks (DSBs). Phosphorylated H2AX has been named γH2AX and its presence in the nucleus can be detected immunocytochemically. Multipara ...

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Detection of DNA Damage Induced by Topoisomerase II Inhibitors, Gamma Radiation and Crosslinking Agents Using the Comet Assay

The comet assay is a simple gel electrophoresis method for visualizing and quantifying DNA damage. The comet assay is sensitive and reproducible and can be used to detect single-strand DNA breaks, double-strand DNA breaks, protein-associated DNA strand breaks and DNA crosslinks. The com ...

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Analysis of Genomic Aberrations Using Comparative Genomic Hybridization of Metaphase Chromosomes

Comparative genomic hybridization (CGH) allows the global screening of copy number aberrations within a sample. Specifically, large (20 mb) deletions and amplifications are detected, which are likely to indicate regions harboring tumor suppressor and oncogenes. CGH involves the ...

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