Abstract:
:Replacement of colorimetric substrates in Western blots by chemiluminescent reagents enhances the sensitivity of detection by more than an order of magnitude. Protein levels beyond the detection limit of colorimetric substrates can be consistently detected without modifying pre-existing laboratory protocols.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Sandhu GS,Eckloff BW,Kline BCsubject
Has Abstractpub_date
1991-07-01 00:00:00pages
14-6issue
1eissn
0736-6205issn
1940-9818journal_volume
11pub_type
杂志文章相关文献
BIOTECHNIQUES文献大全abstract::Address correspondence to Martin A. Kennedy, Department of Pathology & Carney Centre for Pharmacogenomics, University of Otago, Christchurch, PO Box 4345, Christchurch, New Zealand. E-mail: martin.kennedy@otago.ac.nz. ...
journal_title:BioTechniques
pub_type: 信件
doi:10.2144/000114481
更新日期:2016-12-01 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::Procedures utilizing Chelex 100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube transfers for most types of samples. The extraction of DNA from semen and very small blo...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1991-04-01 00:00:00
abstract::Vascular targeting agents (VTAs) can be produced by linking antibodies or antibody fragments directed against endothelial cell markers to effector moieties. So far, it has been necessary to produce the components of VTAs (antibody, antibody fragment, linker, and effector) separately and, subsequently, to conjugate the...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/01301dd03
更新日期:2001-01-01 00:00:00
abstract::We have modified the terminal deoxynucleotidyl transferase (TDT)-mediated dUTP-biotin nick end-labeling (TUNEL) method to permit the immunogold-silver intensification detection of apoptotic cells in tissue sections. Such sections can subsequently be processed for multi-labeling fluorescence microscopy, thus permitting...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1995-11-01 00:00:00
abstract::The use of reverse transcription (RT) PCR for relative quantitation of gene transcripts relies on the reproducibility of the individual RT, PCR and product measurement steps. Semi-competitive RT-PCR (RT-cPCR) uses an internal competitor template in the PCR step to improve quantitation. We have surveyed the reproducibi...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98244rr02
更新日期:1998-04-01 00:00:00
abstract::The degree of fluorescence polarization (FP) of a fluorescent molecule is a reflection of its molecular weight (Mr). FP is therefore a useful detection methodfor homogeneous assays in which the starting reagents and products differ significantly in Mr. We have previously shown that FP is a good detection method for th...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/01313rr01
更新日期:2001-09-01 00:00:00
abstract::Integral membrane G protein-coupled receptors (GPCRs) compose the single most prolific class of drug targets, yet significant functional and structural questions remain unanswered for this superfamily. A primary reason for this gap in understanding arises from the difficulty of forming soluble, monodisperse receptor m...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112169
更新日期:2006-05-01 00:00:00
abstract::We report a modified method of screening for point mutations using a PCR approach based upon the sensitivity of PCR to the 3' terminus of the primer. This method provides a sensitive screen when using either plasmid DNA or bacterial cell lysates. We have optimized the technique for general use to allow rapid screening...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-01-01 00:00:00
abstract::Transfection experiments involving mixed cell cultures pose special problems because of differential uptake and expression of DNA by different cell types. In order to evaluate the expression of transfected DNA in primary cultures of embryonic myocardial cells, we have used immunofluorescence microscopy to analyze expr...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:
更新日期:1988-07-01 00:00:00
abstract::Loop-mediated isothermal amplification (LAMP), a novel gene amplification method, enables the synthesis of larger amounts of both DNA and a visible byproduct--namely, magnesium pyrophosphate--without thermal cycling. A positive reaction is indicated by the turbidity of the reaction solution or the color change after a...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113072
更新日期:2009-03-01 00:00:00
abstract::Reducing the scale of biochemical reactions is becoming commonplace. Examples include the screening of large libraries of chemical compounds or gene sequences. These applications demand the ability to transfer sub-microliter volumes of fluid. To this end, we have modified a Hamilton MICROLAB 2200 with high-speed solen...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/01304rr05
更新日期:2001-04-01 00:00:00
abstract::We describe a simple and efficient system for epitope mapping by cloning random gene fragments into a specially designed gIIIp-based phage display vector. DNA encoding the antigen of interest is PCR-amplified and partially digested with DNaseI to generate 50-150-bp-long fragments, which are polished with T4 DNA polyme...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/99272st04
更新日期:1999-08-01 00:00:00
abstract::A simple, non-destructive procedure is described to determine the quality of DNA arrays before they are used. It consists of a preliminary staining step of the DNA microarray by using SYBR green II, a fluorophore with specific affinity for ssDNA, followed by a laser scan analysis. The surface quality, integrity and ho...
journal_title:BioTechniques
pub_type:
doi:10.2144/00291st01
更新日期:2000-07-01 00:00:00
abstract::Microarrays printed on glass slides are often constructed by covalently linking modified oligonucleotide probes to a derivatized surface at considerable expense. In this article, we demonstrate that 14-base oligonucleotides with a poly(T)10 - poly(C)10 tail (TC tag), but otherwise unmodified, can be linked by UV light...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112905
更新日期:2008-09-01 00:00:00
abstract::Three-dimensional collagen gel contraction is the standard assay utilized for functionally quantifying a variety of cell types, in particular smooth muscle cells (SMCs) and myofibroblasts. Here, we have developed a method to effectively reduce the three-dimensional parameters of the standard collagen gel into a single...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113349
更新日期:2010-02-01 00:00:00
abstract::Enhanced green fluorescent protein (EGFP) is the preferred reporter protein for real-time detection in individual cells, but its usefulness for gene expression quantification is limited by the sensitivity of standard detection techniques. We tested whether the unique feature of single-cell detection and quantification...
journal_title:BioTechniques
pub_type:
doi:10.2144/02321st01
更新日期:2002-01-01 00:00:00
abstract::For a feasible and cost-effective impedance measurement of cellular alterations in real-time, we combined commercially available microelectrode arrays (MEAs), consisting of 60 microelectrodes, with a conventional impedance analyzer. For proof of principle, a breast carcinoma cell line (MCF-7) was cultured on MEAs, and...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112254
更新日期:2006-10-01 00:00:00
abstract::Recombinant retroviral vectors usually encode the genes of interest in place of the viral structural genes, which must be provided in trans. These viruses are therefore defective for replication: infected cells cannot produce progeny virus. However, in some cases it may be desirable to generate virus from an infected ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1994-12-01 00:00:00
abstract::Data are presented illustrating the optimum concentration range of reverse transcription PCR-generated products under 500 bp for accurate base calling with direct automated DNA sequencing. A 357-bp fragment of the human estrogen receptor, which includes the DNA binding domain of the protein, was used as a representati...
journal_title:BioTechniques
pub_type:
doi:
更新日期:1994-09-01 00:00:00
abstract::A new genetic marker was created in which sequences from enhanced green fluorescent protein were fused to those of puromycin N-acetyl transferase. The resulting fusion protein (EGFP-puro) conferred both green fluorescence and resistance to puromycin when expressed in mammalian cells. The utility of EGFP-puro as a sele...
journal_title:BioTechniques
pub_type:
doi:10.2144/01312st05
更新日期:2001-08-01 00:00:00
abstract::The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is rapidly becoming an important reporter molecule for monitoring gene expression and protein localization in vivo, in situ and in real time. GFP emits bright green light (lambda max = 509 nm) when excited with UV or blue light (lambda max = 395 ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1995-10-01 00:00:00
abstract::We describe here a simple and efficient transfection method for transient expression of cloned genes in cell lines and primary cultured cells. The method involves the use of DEAE-dextran to target DNA to the cellular endocytotic pathway and the use of a human adenovirus to ensure efficient lysis of endosomal vesicles....
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1994-08-01 00:00:00
abstract::Normalization is a critical step in the analysis of microarray gene expression data. For dual-labeled array, traditional normalization methods assume that the majority of genes are non-differentially expressed and that the number of overexpressed genes approximately equals the number of under-expressed genes. However,...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113195
更新日期:2009-08-01 00:00:00
abstract::SCORE, a program for computer-assisted scoring of Southern blots of clone DNA, retains the use of expert human judgment while taking over much of the drudgery of the scoring task. The primary functions of the program are to help make an aligned overlay of the fluorescence gel image and the autoradiogram blot image, to...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1991-06-01 00:00:00
abstract::Quantitative PCR assays are now the standard method for viral diagnostics. These assays must be specific, as well as sensitive, to detect the potentially low starting copy number of viral genomic material. We describe a new technique, polymerase chain displacement reaction (PCDR), which uses multiple nested primers in...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113951
更新日期:2013-02-01 00:00:00
abstract::Gene expression analysis using high-density cDNA or oligonucleotide arrays is a rapidly emerging tool for transcriptomics, the analysis of the transcriptional state of a cell or organ. One of the limitations of current methodologies is the requirement of a relatively large amount of total or polyadenylated RNA as star...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/03343rr01
更新日期:2003-03-01 00:00:00
abstract::Exposing cells to brief, high-intensity electrical field pulses can lead to the permeabilization of their plasma membranes. This electro-induced permeated state of the cell membrane is reversible and leads to cell fusion; i.e., the electropermeabilized state if fusogenic. The size of cells intended for fusing, however...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1994-10-01 00:00:00
abstract::Reverse transcription PCR (RT-PCR) represents a sensitive and powerful tool for analyzing RNA. While it has tremendous potential for quantitative applications, a comprehensive knowledge of its technical aspects is required. Successful quantitative RT-PCR involves correction for experimental variations in individual RT...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:10.2144/99261rv01
更新日期:1999-01-01 00:00:00
abstract::Perfluorocarbon emulsions were applied to hybridoma cultures grown in tissue culture tubes and column bioreactors. The oxygen transfer enhancement effect of perfluorocarbon emulsions was clearly demonstrated by the higher cell densities obtained in emulsion-supplemented systems. In addition, perfluorocarbon emulsions ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-02-01 00:00:00