Double staining to increase the sensitivity of protein detection in polyacrylamide gels.

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

authors： Morseman JP,Moss MW,Zoha SJ,Allnutt FC

更新日期：1999-03-01 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

authors： Yang XP,Gallo M,Ngan I,Nocerini M,Chen MM

更新日期：2002-03-01 00:00:00

abstract：:In the design of oligonucleotide sequences for targeting DNA or RNA sequences, it can be difficult to identify sequences that will hybridize only to the intended target. The term "sequence-specific" or "sequence-nonspecific" is often used to describe the interactions of an oligonucleotide with a mixture of DNA or RNA....

journal_title：BioTechniques

authors： Hyndman D,Cooper A,Pruzinsky S,Coad D,Mitsuhashi M

更新日期：1996-06-01 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

authors： Corbley MJ,Cherington V,Feig LA,Cooper GM,Roberts TM

更新日期：1994-12-01 00:00:00

abstract：:Characterization of the thermodynamics of binding interactions is important in improving our understanding of bimolecular recognition and forms an essential part of the rational drug design process. Isothermal titration calorimetry (ITC) is rapidly becoming established as the method of choice for undertaking such stud...

journal_title：BioTechniques

authors： Holdgate GA

更新日期：2001-07-01 00:00:00

abstract：:DNA damage in the form of abasic sites, chemically altered nucleotides, and strand fragmentation is the foremost limitation in obtaining genetic information from many ancient samples. Upon cell death, DNA continues to endure various chemical attacks such as hydrolysis and oxidation, but repair pathways found in vivo n...

journal_title：BioTechniques

authors： Mouttham N,Klunk J,Kuch M,Fourney R,Poinar H

更新日期：2015-07-01 00:00:00

abstract：:A 200-year-old observation might provide a new way to eliminate tumors by infecting cancer patients with bacteria. Kristie Nybo explores a new approach that could transform cancer treatment. ...

journal_title：BioTechniques

authors： Nybo K

更新日期：2018-01-01 00:00:00

abstract：:Pairs of primers flanking known miniTn10 transposon insertion sites were used to confirm the presence of the transposon in DNA isolated from Legionella pneumophila mutants. It was expected that the polymerase chain reaction products derived from the mutant template would be larger than those from the wild-type (WT) te...

journal_title：BioTechniques

authors： Viswanathan VK,Krcmarik K,Cianciotto NP

更新日期：1999-09-01 00:00:00

abstract：:Complete brain fixation can be achieved with transthoracic cardiac infusion without thoracotomy. Light and electron microscopy tissue sections reveal preservation of cytoplasmic and nuclear structure at all magnification levels. Punched samples were obtained from the fixed tissue specimens in precisely localized areas...

journal_title：BioTechniques

authors： Eichenbaum KD,Eichenbaum JW,Fadiel A,Miller DC,Demir N,Naftolin F,Stern A,Pevsner PH

更新日期：2005-10-01 00:00:00

abstract：:Single-stranded long oligonucleotide-based (50- to 70-mer) microarrays offer several advantages over conventional cDNA microarrays. These include the easy preparation of the probes, low cost of array production, and low cross-contamination during probe handling. However, the application of oligonucleotide microarrays ...

journal_title：BioTechniques

authors： Kaposi-Novak P,Lee JS,Mikaelyan A,Patel V,Thorgeirsson SS

更新日期：2004-10-01 00:00:00

abstract：:Cell-imaging approaches using new laser-based technologies have a wide applicability to thefields of pathology and cell biology. Here, we present the application of several of these techniques, including confocal scanning laser microscopy (CSLM), laser scanning cytometry (LSC), and laser capture microdissection (LCM),...

journal_title：BioTechniques

authors： Taatjes DJ,Palmer CJ,Pantano C,Hoffmann SB,Cummins A,Mossman BT

更新日期：2001-10-01 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

authors： Qureshi SA

更新日期：2007-01-01 00:00:00

abstract：:Cytokines are pivotal to a balanced innate or cell-mediated immune response, can be indicative of disease progression and/or resolution, and are being evaluated as therapeutics. There is a need to purify and/or to measure key cytokines rapidly with accuracy, precision, and sensitivity. The current assay technologies, ...

journal_title：BioTechniques

authors： Bai H,Buller RM,Chen N,Boyle MD

更新日期：2002-01-01 00:00:00

abstract：:The conventional method for cloning a DNA fragment is to insert it into a vector and ligate it. Although this method is commonly used, it is labor intensive because the ratio and concentrations of the DNA insert and the vector need optimizing. Even then, the resultant library is often plagued with unwanted plasmids th...

journal_title：BioTechniques

authors： Miyazaki K,Takenouchi M

更新日期：2002-11-01 00:00:00

abstract：:High affinity aptamer-based biomarker discovery has the advantage of simultaneously discovering an aptamer affinity reagent and its target biomarker protein. Here, we demonstrate a morphology-based tissue aptamer selection method that enables us to use tissue sections from individual patients and identify high-affinit...

journal_title：BioTechniques

authors： Wang H,Li X,Volk DE,Lokesh GL,Elizondo-Riojas MA,Li L,Nick AM,Sood AK,Rosenblatt KP,Gorenstein DG

更新日期：2016-11-01 00:00:00

abstract：:We present a tool for dispensing very low volumes (20 nL or more) of ultra high viscosity (UHV) medical-grade alginate hydrogels. It uses a modified piezo-driven micrometering valve, integrated into a versatile system that allows fast prototyping of encapsulation procedures and scaffold production. Valves show excelle...

journal_title：BioTechniques

authors： Gepp MM,Ehrhart F,Shirley SG,Howitz S,Zimmermann H

更新日期：2009-01-01 00:00:00

abstract：:We describe the application of simple cloning by prolonged overlap extension for multiple site-directed mutagenesis in the same plasmid. We show that it is possible to use this technique with very short PCR templates. The technique is ideally suited for the generation of longer donor DNA sequences for CRISPR/Cas9-medi...

journal_title：BioTechniques

authors： Hejlesen R,Füchtbauer EM

更新日期：2020-06-01 00:00:00

abstract：:Bovine serum albumin, free of deoxyribonuclease activity, was obtained in our laboratory using ion-exchange chromatography followed by acetylation. Chromatography on four different resins (DEAE-52, P-11, hydroxylapatite and Q Sepharose fast-flow) was examined. Fractions from Q Sepharose chromatography, eluted with a l...

journal_title：BioTechniques

authors： Trujillo LE,Castellanos L,Garcia O,Herrera L

更新日期：1990-11-01 00:00:00

abstract：:A brief description of the setup of an inexpensive densitometric analysis system is provided. This system uses a software package and scanner that can be obtained for a total of less than $1000 and that can be operated by any Macintosh computer, including a MacPlus. In the event that a scanner is already available, th...

journal_title：BioTechniques

authors： Shea TB

更新日期：1994-06-01 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

authors： Leitz AJ,Bayburt TH,Barnakov AN,Springer BA,Sligar SG

更新日期：2006-05-01 00:00:00

abstract：:We observed the presence of contaminating NADH oxidation activity in maltose binding protein (MBP) fusion proteins expressed in Escherichia coli and purified using conventional amylose resin-based affinity chromatography. This contaminating NADH oxidation activity was detectable with at least four different enzymes fr...

journal_title：BioTechniques

authors： Guo F,Zhu G

更新日期：2012-04-01 00:00:00

abstract：:The increase of information in biology makes it difficult for researchers in any field to keep current with the literature. The MEDLINE database of scientific abstracts can be quickly scanned using electronic mechanisms. Potentially interesting abstracts can be selected by matching words joined by Boolean operators. H...

journal_title：BioTechniques

authors： Perez-Iratxeta C,Keer HS,Bork P,Andrade MA

更新日期：2002-06-01 00:00:00

abstract：:A nascent protein remains in a complex with its ribosome and mRNA if the stop codon is deleted from the mRNA. In the same manner, mRNA forms a stable complex with DNA if the transcription termination is blocked. In principle, if both mRNA translation and DNA transcription termination are prevented, the protein should ...

journal_title：BioTechniques

authors： Liu S,Shivashankar GV,Sano T,Libchaber A

更新日期：2000-10-01 00:00:00

abstract：:Here we describe a method for the isolation of PCR-ready genomic DNA from various zebrafish tissues that is based on a previously published murine protocol. The DNA solutions are of sufficient quality to allow PCR detection of transgenes from all commonly used zebrafish tissues. In sperm, transgene amplification was s...

journal_title：BioTechniques

authors： Meeker ND,Hutchinson SA,Ho L,Trede NS

更新日期：2007-11-01 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：:This report describes a common method of obtaining template DNA from phagemids, phages and plasmids. The strategy is based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB) for DNA precipitation. By avoiding phase separation, many manipulation steps are reduced. A time-saving modification to ...

journal_title：BioTechniques

authors： Del Sal G,Manfioletti G,Schneider C

更新日期：1989-05-01 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 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：: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 resolution of complex protein mixtures by discontinuous buffer SDS-PAGE is accomplished by their concentration into thin bands in the stacking gel, followed by their separation during migration through the resolving gel. Recombinant human interferon-inducible protein-10 (IP-10), a 10-kDa C-X-C chemokine with four ...

journal_title：BioTechniques

authors： Crow MK,Karasavvas N,Sarris AH

更新日期：2001-02-01 00:00:00