Cold-adapted protease enables quantitation of surface proteins in the absence of membrane trafficking.

abstract：:A method for the determination of prostaglandin G/H synthase and lipoxygenase activities in tissues was developed and employed with rat gastric mucosa samples. Tissues and microsomes were incubated in a buffer containing nonionic detergent and 1.32 mM arachidonic acid for 10 min. Following extraction with ethyl acetat...

journal_title：BioTechniques

authors： Stein TA,Bailey B,Auguste LJ,Wise L

更新日期：1991-02-01 00:00:00

abstract：:Human embryonic kidney (HEK293) cells were stably transduced with a retroviral vector containing an expression cassette for a short-lived green fluorescent protein (d2EGFP) and the neomycin resistance gene (Neor). When Neor HEK293 clones were treated with proteasome inhibitors, lactacystin or MG132, an increase in the...

journal_title：BioTechniques

authors： Andreatta C,Nahreini P,Hovland AR,Kumar B,Edwards-Prasad J,Prasad KN

更新日期：2001-03-01 00:00:00

abstract：:We have developed a high-throughput, multiplex reverse transcription PCR (RTPCR) assay that is suitable for the analysis of medium-to low-copy cellular RNA transcripts from small numbers of cells (10(4)). High throughput was attained by utilizing microplate-based RNA extraction and RTPCR protocols, followed by PCR pro...

journal_title：BioTechniques

authors： Su S,Vivier RG,Dickson MC,Thomas N,Kendrick MK,Williamson NM,Anson JG,Houston JG,Craig FF

更新日期：1997-06-01 00:00:00

abstract：:The serum protein transferrin (Tf) is a valuable marker for genetic studies of primates, because it is usually polymorphic, exhibiting as many as 13 allelic forms with high heterozygosity. The standard procedure to detect the different phenotypes requires vertical electrophoresis on polyacrylamide gels for 18 h at 4 d...

journal_title：BioTechniques

authors： Manis GS,Samples NK,Stone WH

更新日期：1993-11-01 00:00:00

abstract：:Ribonucleases H (RNases H) are enzymes that specifically degrade the RNA of RNA-DNA hybrids. These enzymes are involved in DNA replication, reverse transcription (RT) and antisense oligodeoxyribonucleotide-mediated arrest of translation. One of the most valuable tools for assaying RNase H activity is the renaturation ...

journal_title：BioTechniques

authors： Han LY,Ma WP,Crouch RJ

更新日期：1997-11-01 00:00:00

abstract：:Plasmid DNA can be purified in 20 min using a three-step CsCl gradient in a vertical tube rotor. Ultracentrifugation with discontinuous gradients yields DNA with purity comparable to that obtained with longer isopycnic separations and with columns. ...

journal_title：BioTechniques

authors： Dorin M,Bornecque CA

更新日期：1995-01-01 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

authors： Gorski DH,LePage DF,Walsh K

更新日期：1994-05-01 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

authors： Ross M,Peters L

更新日期：1990-11-01 00:00:00

abstract：:RNA interference (RNAi) is a recent advance that provides the possibility to reduce the expression of specific target genes in cultured mammalian cells with potential applications on a genome-wide scale. However, to achieve this, robust methodologies that allow automated and efficient delivery of small interfering RNA...

journal_title：BioTechniques

authors： Erfle H,Simpson JC,Bastiaens PI,Pepperkok R

更新日期：2004-09-01 00:00:00

abstract：:Expanding applications of cDNA microarrays such as fine needle aspiration biopsy and laser capture microdissection necessitate the ability to perform arrays with minute starting amounts of RNA. While methods for amplifying RNA have been advocated, the fidelity of array results using amplified material has not been ful...

journal_title：BioTechniques

authors： Feldman AL,Costouros NG,Wang E,Qian M,Marincola FM,Alexander HR,Libutti SK

更新日期：2002-10-01 00:00:00

abstract：:The microcomputer-based image analysis system IB-1000 (developed by Indiana Biotech, Highland, IN) for two-dimensional electrophoresis gels has been described previously (9). It allows the user to compare protein spots between two profiles and identify those spots that are commonly shared in both profiles. This report...

journal_title：BioTechniques

authors： Lee C,Sherwood ER,Sensibar JA,Berg LA,Chen YC,Tseng CC

更新日期：1989-04-01 00:00:00

abstract：:Current protocols for screening proliferative factors for β cells ex vivo are time-consuming, require cell lines or dissociated islets, and often entail expensive specialized screening equipment. Here we present an efficient and lower cost alternative that utilizes intact mouse islets for the initial screening of prol...

journal_title：BioTechniques

authors： Mosser RE,Gannon M

更新日期：2013-12-01 00:00:00

abstract：:Based on the requirement of a glutathione-deficient mutant strain of Saccharomyces cerevisiae to take up external glutathione for growth on synthetic media, a simple agar diffusion test for quantitative detection of total glutathione from various sources was established. Glutathione concentrations can be reliably dete...

journal_title：BioTechniques

authors： Schmidt M,Grey M,Brendel M

更新日期：1996-11-01 00:00:00

abstract：:Replication studies on human immunodeficiency virus 1 (HIV-1) rely on a few laboratory strains that are divergent from dominant HIV-1 subtypes in the epidemic. Several phenotypic differences between diverse HIV-1 isolates and subtypes could affect vaccine development and treatment, but this research field lacks robust...

journal_title：BioTechniques

authors： Dudley DM,Gao Y,Nelson KN,Henry KR,Nankya I,Gibson RM,Arts EJ

更新日期：2009-05-01 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

authors： Zhao S,Ooi SL,Pardee AB

更新日期：1995-05-01 00:00:00

abstract：:Radiolabeling of native proteins conventionally has required iodination using 125Iodine (125I). Although radioiodination can result in high specific activity, there are several drawbacks in the use of 125I (e.g., radiological hazards and short half-life). 14C-Methylamine-glutaraldehyde conjugation to proteins offers a...

journal_title：BioTechniques

authors： Moore PL,Damelin LH,Harrison TJ

更新日期：2003-08-01 00:00:00

abstract：:The construction and the testing of two lambda phage vectors are described that greatly simplify the tasks of mapping genomic DNA and making replacement-type gene-targeting vectors for mammalian cells from a library of isogenic genomic DNA. The first vector, lambda PS, accommodates up to 20 kb and allows inserts to be...

journal_title：BioTechniques

authors： Nehls M,Messerle M,Sirulnik A,Smith AJ,Boehm T

更新日期：1994-10-01 00:00:00

abstract：:We have utilized 5-bromo-2'deoxyuridine (BrdU) substituted DNA as a probe for a number of applications including, principally, for chromosome painting by fluorescence in situ hybridization (FISH) but also for DNA end-labeling to detect apoptotic cell death and for filter hybridization. Br-dUTP was used as a substitute...

journal_title：BioTechniques

authors： Mühlmann-Díaz MC,Dullea RG,Bedford JS

更新日期：1996-07-01 00:00:00

abstract：:3D printed biomaterials are increasingly used in cell cultures and drug screens. Given the ease of creating artificial tissues, will this technique revolutionize biomedicine and organ implants in the future? ...

journal_title：BioTechniques

authors： Prabhune M

更新日期：2018-03-01 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

authors： Elkin C,Kapur H,Smith T,Humphries D,Pollard M,Hammon N,Hawkins T

更新日期：2002-06-01 00:00:00

abstract：:Several DNA extraction methods have been reported for use with digesta or fecal samples, but problems are often encountered in terms of relatively low DNA yields and/or recovering DNA free of inhibitory substances. Here we report a modified method to extract PCR-quality microbial community DNA from these types of samp...

journal_title：BioTechniques

authors： Yu Z,Morrison M

更新日期：2004-05-01 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

authors： Heda GD,Shrestha L,Thapa S,Ghimire S,Raut D

更新日期：2020-06-01 00:00:00

abstract：:A simple and rapid method for permanently marking autoradiographs is described. This procedure is based on the phosphorescence of light-activated zinc sulfide and the subsequent exposure of x-ray films by this light emission. A lacquer-based carrier allows the zinc sulfide to remain in suspension and permits permanent...

journal_title：BioTechniques

authors： Seto D,Rohrabacher C

更新日期：1991-05-01 00:00:00

abstract：:As a strategy to purify recombinant murine Interleukin (IL)-18, we cloned the mature coding region of this protein into the pFLAG-1 expression system. The intent was to use the FLAG peptide "tag" as an amino terminal addition to IL-18 so that purification of this fusion protein (FLAG-IL-18) on anti-FLAG antibody affin...

journal_title：BioTechniques

authors： Elhofy A,Bost KL

更新日期：1998-09-01 00:00:00

abstract：:The Drosophila heart has gained considerable traction as a model of cardiac development and physiology. Previously we described a semiautomatic optical heartbeat analysis (SOHA) method for quantifying functional parameters from the fly heart that facilitated its use as an organ system and disease model. Here we presen...

journal_title：BioTechniques

authors： Cammarato A,Ocorr S,Ocorr K

更新日期：2015-02-01 00:00:00

abstract：:A new qualitative PCR product detection assay called competitive amplified single mutation detection by selective probe hybridization immunoassay (CASSI) was developed for genotyping the most common apolipoprotein E (apoE) polymorphisms. Single target DNA strands immobilized using biotin on streptavidin-coated micropl...

journal_title：BioTechniques

authors： Köhler T,Rost AK,Purschwitz K,Vondran S,Remke H,Wagner O,Richter V

更新日期：1998-07-01 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

authors： Hegde P,Qi R,Abernathy K,Gay C,Dharap S,Gaspard R,Hughes JE,Snesrud E,Lee N,Quackenbush J

更新日期：2000-09-01 00:00:00

abstract：:14CO2 capture is commonly used to evaluate the cellular oxidation rate of respiratory substrates. A modification of the established 14CO2-capture method was developed that enables the use of cells in adherent culture and easy analysis of multiple samples under different culture conditions. The use of commercially avai...

journal_title：BioTechniques

authors： Collins CL,Bode BP,Souba WW,Abcouwer SF

更新日期：1998-05-01 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

authors： Harris CL,Sanchez-Vargas IJ,Olson KE,Alphey L,Fu G

更新日期：2013-02-01 00:00:00

abstract：:Sample automation and management is increasingly important as the number and size of population-scale and high-throughput projects grow. This is particularly the case in large-scale population studies where sample size is far outpacing the commonly used 96-well plate format. To facilitate management and transfer of sa...

journal_title：BioTechniques

authors： Cario CL,Witte JS

更新日期：2018-12-01 00:00:00