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Purification of Recombinant Proteins from E. coli by Engineered Inteins

The IMPACT (Intein-Mediated Purification with an Affinity Chitin-Binding Tag) vectors are designed for the isolation of pure, functional, recombinant proteins by a single affinity chromatography step. The IMPACT technology was developed at New England Biolabs (NEB) by exploiti ...

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Calmodulin as an Affinity Purification Tag

Calmodulin is a small (148 amino acids, 17 kDa) ubiquitous protein in eukaryotes, and is considered the primary intracellular calcium sensor, making it a key regulator of intracellular signal transduction. Upon calcium binding, calmodulin can interact with a variety of proteins (1), medi ...

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Maltose-Binding Protein as a Solubility Enhancer

A major impediment to the production of recombinant proteins in Escherichia coli is their tendency to accumulate in the form of insoluble and biologically inactive aggregates known as inclusion bodies. Although it is sometimes possible to convert aggregated material into native, bio ...

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Calmodulin-Binding Peptide as a Removable Affinity Tag for Protein Purification

Protein purification is an important tool for investigations on protein function, structure analysis, and biotechnological use. Therefore a number of different techniques have been developed for fast, reliable, and reproducible overexpression and purification of relevant ...

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Thioredoxin and Related Proteins as Multifunctional Fusion Tags for Soluble Expression in E. coli

Escherichia coli has traditionally been a popular host for the production of heterologous proteins because of its ease of genetic manipulation and growth. Recombinant proteins produced in E. coli have been useful for a variety of purposes, including the study of protein tertiary structu ...

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Discovery of New Fusion Protein Systems Designed to Enhance Solubility in E. coli

Fusion protein technology has been creatively applied to solve many problems encountered in the study of protein structure and function (1,2). One prevalent application is the use of fusion proteins to improve protein expression in Escherichia coli and provide convenient methods for a ...

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Detection of Anti-GOR Antibodies in HCV Infection

The interactions of the hepatitis C virus (HCV) with the immune system are numerous. As one of the many results, a considerable number of autoantibodies occur in serum. This chapter describes methods to detect anti-GOR antibodies (1–3). Anti-GOR are autoantibodies that are specifically ass ...

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The Chimpanzee Model: Contributions and Considerations Elizabeth Muchmore

Chimpanzees (Pan troglodytes) became established as invaluable models for the study of human viral hepatitis after it was discovered, in 1967, that the chronic hepatitis B antigen carrier state existed in a naturally infected member of this species (1–3). They were instrumental in the devel ...

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Confirmation of HCV Antibodies by the Line Immunoassay INNO-LIA HCV Ab III

HCVs constitute a genus within the Flaviviridae, with closest homology to the hepatitis G and GB viruses, and Pestiviruses. The positive-stranded RNA genome encodes at least nine proteins. Core, El, and E2 constitute the structural proteins; NS2, NS3, NS4A, NS4B, NS5A, and NS5B are nonstructur ...

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Liver Transplantation as a Model to Study Hepatitis C Virus Infection

Hepatitis C virus-associated end-stage liver disease is a leading diagnosis in patients undergoing liver transplantation, accounting for approx 25% of patients transplanted at major medical centers in the United States, and for 5–15% of those transplanted worldwide (1). Although HCV ...

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Detection of HCV RNA in Serum by Reverse Transcriptase-PCR and Radiolabeled Liquid Hybridization

Hepatitis C virus (HCV) possesses a single-stranded, positive-sense RNA that is 9.4 kb in length. The complete HCV genome has been cloned and sequenced and encodes for a nucleocapsid, an envelope, and five nonstructural proteins (1,2). The 5′ untranslated region of the virus is highly conserved am ...

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The AMPLICOR HCV Tests for the Detection and Quantitation of Serum or Plasma HCV RNA

Clinical diagnosis of HCV infection is generally accomplished by using immunoserological assays to detect the presence of anti-HCV antibodies. Such immunoserological assays have been approved for blood donor screening, thereby reducing the incidence of post-transfusion he ...

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Detection of HCV RNA in Serum by Reverse Transcription Polymerase Chain Reaction (RT-PCR)

In the field of biological science, the development of new techniques (e.g., Southern blotting, molecular cloning, pulsed-field gel electrophoresis) often led to better understanding of fundamental and applied biological problems. The polymerase chain reaction (PCR) is among the t ...

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Quantification of HCV RNA in Clinical Specimens by Branched DNA (bDNA) Technology

The diagnosis and monitoring of hepatitis C virus (HCV) infection have been aided by the development of HCV RNA quantification assays A direct measure of viral load, HCV RNA quantification has the advantage of providing information on viral kinetics and provides unique insight into the dise ...

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Quality Control of the Polymerase Chain Reaction

The polymerase chain reaction (PCR) has revolutionized both the basic and the applied aspects of the biomedical field with more than 40,000 papers having been published employing this technique. PCR has become an indispensable tool for basic research applications, such as cloning (1), seq ...

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Detection of HCV RNA in Formalin-Fixed, Paraffin-Embedded Liver Tissue by RT-PCR

Hepatitis C virus (HCV) RNA has been detected in the sera and liver sections of patients with chronic HCV infection using RT-PCR and other sensitive molecular techniques. Since archived formalin-fixed, paraffin embedded tissues from these patients are readily available for retrospe ...

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Quantification of HCV RNA in Liver Tissue by bDNA Assay

With this statement, Sherlock and Dooley have described two of the three major challenges involved in quantitatively measuring any analyte in tissue samples: the distribution of the analyte in the tissue; and the standard of reference, or denominator, with which to make comparisons betwe ...

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Preparation of Genotype-Specific HCV RNA Transcripts for Assessing HCV Detection and Quantification Assays

Hepatitis C virus (HCV), the etiological agent responsible for the majority of cases of parenterally acquired liver disease, is found throughout the world. HCV is an enveloped virus with a small, single-stranded RNA genome. Because it uses an error-prone, RNA-dependent RNA polymerase, HCV h ...

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Hepatitis C Virus: Types, Subtypes, and Beyond

Non-A, non-B hepatitis was recognized as a frequent consequence of blood transfusion for many years before the agent responsible, hepatitis C virus (HCV), was first cloned and sequenced in 1989 Very quickly it became apparent that viruses from different parts of the world were distinct, and aft ...

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Molecular Evolutionary Analysis: Its Application in the Study of Hepatitis C Virus

The recent development of high-power personal computer hardware and software has allowed investigators to prepare phylogenetic trees without understanding of the fundamental principles of molecular evolution. Unfortunately, inexperienced investigators are also pr ...

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