Abstract：
:The use of affinity-based tools has become invaluable as a platform for basic research and in the development of drugs and diagnostics. Applications include affinity chromatography and affinity tag fusions for efficient purification of proteins as well as methods to probe the protein network interactions on a whole-proteome level. A variety of selection systems has been described for in vitro evolution of affinity reagents using combinatorial libraries, which make it possible to create high-affinity reagents to virtually all biomolecules, as exemplified by generation of therapeutic antibodies and new protein scaffold binders. The strategies for high-throughput generation of affinity reagents have also opened up the possibility of generating specific protein probes on a whole-proteome level. Recently, such affinity proteomics have allowed the detailed analysis of human protein expression in a comprehensive manner both in normal and disease tissue using tissue microarrays and confocal microscopy.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Uhlén Mdoi
10.2144/000112803subject
Has Abstractpub_date
2008-04-01 00:00:00pages
649-54issue
5eissn
0736-6205issn
1940-9818pii
000112803journal_volume
44pub_type
杂志文章,评审相关文献
BIOTECHNIQUES文献大全abstract：:Hereditary hemochromatosis (HHC) represents an autosomal recessive disease in which increased iron absorption causes iron overload and irreversible tissue damage. The recently detected association between two point mutations in the HFE gene on chromosome 6p and HHC has made it possible to screen for the disease before...
journal_title：BioTechniques
pub_type：
doi：10.2144/99266st02
更新日期：1999-06-01 00:00:00
abstract：:Tissue engineering scaffolds have emerged as a powerful tool within regenerative medicine. These materials are being designed to create environments that promote regeneration through a combination of: (i) scaffold architecture, (ii) the use of scaffolds as vehicles for transplanting progenitor cells, and/or (iii) loca...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：10.2144/000113754
更新日期：2011-10-01 00:00:00
abstract：:The use of quantitative PCR is recommended to monitor the level of residual hematological malignancies. The proposed multiplex IgH/ras PCR uses a co-amplification of the clonal CDR3 rearrangement of the immunoglobulin heavy chain gene (IgH) as a disease marker and a segment of the Hras 1 gene containing codon 61 (ras)...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/00284st08
更新日期：2000-04-01 00:00:00
abstract：:Random mutagenesis followed by an in vitro selection procedure was shown to be capable of identifying important bases of the hairpin ribozyme for cleavage of an RNA target sequence. The selection scheme enriched the RNA population for those molecules capable of efficient site-specific self-cleavage in the absence of l...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/98242st05
更新日期：1998-02-01 00:00:00
abstract：:For immunological research on the human cytomegalovirus (HCMV), a virus that combines the broad cell tropism of clinical isolates, efficient replication in cell culture, the complete set of MHC-I modulator genes, and suitability for genetic engineering is desired. Here, we aimed to generate a genetically complete deri...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114606
更新日期：2017-11-01 00:00:00
abstract：:Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a ...
journal_title：BioTechniques
pub_type：
doi：10.2144/02323dd05
更新日期：2002-03-01 00:00:00
abstract：:As more and more researchers are examining proteins that are available only in extremely limited quantities, i.e., cellular extracts or genetic engineering products, it is critical to utilize staining methods that maximize sensitivity. The protocol we describe here--double staining of polyacrylamide electrophoresis ge...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1990-11-01 00:00:00
abstract：:SNPCEQer identifies and reports SNPs in sequences obtained from the Beckman CEQ2000 DNA Analysis System. SNPCEQer aligns sequences obtained using CEQ2000 heterozygote detection analysis and reports discrepancies between individual sequences and the consensus sequence it generates from this set as SNPs when the individ...
journal_title：BioTechniques
pub_type：
doi：10.2144/02334st05
更新日期：2002-10-01 00:00:00
abstract：:An improved Huffman coding method for information storage in DNA is described. The method entails the utilization of modified unambiguous base assignment that enables efficient coding of characters. A plasmid-based library with efficient and reliable information retrieval and assembly with uniquely designed primers is...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000113218
更新日期：2009-09-01 00:00:00
abstract：:A fully automated nucleic acid analysis system is described, which offers positive sample identification, improved sensitivity and reduced user interaction compared to conventional techniques. The system relies on the sequence-specific capture of DNA onto solid-phase particles, confirming product identity without the ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/99271pf01
更新日期：1999-07-01 00:00:00
abstract：:An easy, sensitive and direct fluorescent immunodetection method for proteins is described using the new fluorochrome PBXL-1 imaged with the FMBIO II Laser Scanning Imaging System. PBXL-1 is derived from a protein supra-molecular complex that contains a large number of chromophores. This complex, the phycobilisome, is...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/99263pf02
更新日期：1999-03-01 00:00:00
abstract：:Much effort has been directed toward the isolation and characterization of homeobox cDNAs from numerous cell types because they encode transcription factors important to many cellular processes, including pattern formation in the embryo, cell growth and cell differentiation. Many novel homeobox cDNAs have been isolate...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-05-01 00:00:00
abstract：:We describe a rapid, simple and inexpensive method for the isolation of DNA from blood clots suited for use in PCR. Our method is based on the lysing and nuclease-inactivating properties of guanidine thiocyanate together with the nucleic acid-binding properties of silica particles. Isolated DNA can be used for in vitr...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1993-02-01 00:00:00
abstract：:The recently developed ultrasensitive reverse transcriptase (RT) test involving a PCR step can detect minute amounts of RT in single retroviral particles and is 10(6) times more sensitive than conventional RT assays. We have found that different DNA-dependent DNA polymerases like DNA Polymerase I from Escherichia coli...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1996-02-01 00:00:00
abstract：:Here we introduce a modified antibody staining method that uses up to 80% less antibody for flow cytometry. We demonstrate this method for the detection of antigens expressed at high, moderate, or low levels in mouse and rat lymphocytes as well as mouse mammary epithelial cells. We obtained reproducibly accurate resul...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/0000113854
更新日期：2012-07-01 00:00:00
abstract：:We have developed an automated purification method for dye-terminator-based DNA sequencing products using a magnetic bead approach. This 384-well protocol generates sequence fragments that are essentially free of template DNA, salt, and excess dye-terminator products. In comparison with traditional ethanol precipitati...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/02326st05
更新日期：2002-06-01 00:00:00
abstract：:The Cre/lox system is a powerful genetic tool with which to manipulate the genome. Here, we describe the development of a simple reporter system for Cre recombinase, called the Cre Stoplight. In the absence of Cre, the red fluorescent protein is expressed; when Cre catalyzes a recombination event, the green fluorescen...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/01315st03
更新日期：2001-11-01 00:00:00
abstract：:Microarray expression analysis has become one of the most widely used functional genomics tools. Efficient application of this technique requires the development of robust and reproducible protocols. We have optimized all aspects of the process, including PCR amplification of target cDNA clones, microarray printing, p...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：10.2144/00293bi01
更新日期：2000-09-01 00:00:00
abstract：:The ZEBRA protein is a transcriptional activator that induces expression of viral lytic genes in cells harboring latent Epstein-Barr virus (EBV). In this report it is shown that a derivative of ZEBRA that cannot activate transcription (Zd) can be used to detect and characterize activation domains. Three expression vec...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/97225rr02
更新日期：1997-05-01 00:00:00
abstract：:We describe a new real-time fluorescence video microscope design for capturing intensified images of cells containing dual wavelength "ratio" dyes or multiple dyes. The microscope will perform real-time capture of two separate fluorescence emission images simultaneously, improving the time resolution of spatial distri...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1990-03-01 00:00:00
abstract：:To gain insightful information about the mechanisms through which genes are activated and repressed requires gene reporter systems that are sensitive, robust, and cost-effective. Although numerous reporter gene technologies are commercially available, none are as sophisticated and user-friendly as beta-lactamase (BLA)...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：10.2144/000112292
更新日期：2007-01-01 00:00:00
abstract：:The two-hybrid system is a genetic method to identify proteins that interact with a specific target protein, which is expressed in yeast as a hybrid with a DNA-binding domain. Use of this method entails transforming yeast, both with this DNA-binding domain hybrid and with a library of activation domain hybrids, follow...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1993-06-01 00:00:00
abstract：:Radiolabeled proteins are used in a variety of laboratory applications as well as in radioimmunotherapy. This review focuses on methods that utilize genetic engineering to introduce exogenous phosphorylation sites into proteins. Protein kinase substrate sites can be introduced into target proteins to serve as tags for...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：
更新日期：2002-10-01 00:00:00
abstract：:We developed a sensitive fluorescence assay for the quantitation of proteins in solution using the NanoOrange reagent, a merocyanine dye that produces a large increase in fluorescence quantum yield upon interaction with detergent-coated proteins. The NanoOrange assay allowed for the detection of 10 ng/mL to 10 microgr...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/03344pt03
更新日期：2003-04-01 00:00:00
abstract：:We have developed a novel method for the rapid preparation of large quantities of 3' end-labeled single-stranded (ss) DNA (ssDNA) probes for transcript mapping. A recombinant phagemid vector containing the probe sequence was used to raise large quantities of ssDNA. Based on the DNA sequence of the probe, an oligonucle...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1993-05-01 00:00:00
abstract：:Current gene synthesis methods often incorporate a PCR amplification step in order to yield final material sufficient for resolution from multiple off-products. These amplification steps can cause stochastic sampling effects that propagate errors in gene synthesis or decrease variability when applied to the constructi...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114330
更新日期：2015-09-01 00:00:00
abstract：:Quantitative real-time PCR has become a popular method to analyze and quantify changes in the copy number of mitochondrial DNA (mtDNA), and nuclear DNA (nDNA) is often used as an endogenous reference for mtDNA abundance. In our experience, using nDNA as a reference is problematic, due to differences in the extraction ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000113222
更新日期：2009-10-01 00:00:00
abstract：:Protein samples electroblotted onto nitrocellulose membranes and quenched with a mixture of blocking agents produced a strong signal for cystic fibrosis transmembrane-conductance regulator (CFTR), a high-molecular-weight protein, in western blotting. Optimized conditions for CFTR were then extended to medium- and low-...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/btn-2019-0124
更新日期：2020-06-01 00:00:00
abstract：:To increase the reproducibility and to reduce the false positives in the initial mRNA differential display, modified long composite primers were developed based on both mRNA differential display and RNA arbitrarily primed PCR fingerprinting methods. Ten-base nucleotides were added at the 5' ends of the primers used in...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1995-05-01 00:00:00
abstract：:Site-specific mutagenesis and directional subcloning were accomplished by using the polymerase chain reaction to generate products that can recombine to form circular DNA. This DNA was transfected into E. coli without phosphorylation of primers, restriction enzyme digestion or ligation. Specifically, the polymerase ch...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1990-02-01 00:00:00
