Site-specific 32P-labeling of cytokines, monoclonal antibodies, and other protein substrates for quantitative assays and therapeutic application.

abstract：:The role model systems have played in understanding telomere biology has been enormous, and understanding has rapidly transferred to human telomere research. Most work using model organisms to study telomerase and nontelomerase-based telomere-maintenance systems has centered on yeasts, ciliates, and insects. But it is...

journal_title：BioTechniques

authors： Fajkus J,Sýkorová E,Leitch AR

更新日期：2005-02-01 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

authors： Scherer A,Krause A,Walker JR,Sutton SE,Serón D,Raulf F,Cooke MP

更新日期：2003-03-01 00:00:00

abstract：:Stable cell transfection is used for the expression of exogenous genes or cDNAs in eukaryotic cells. Selection of these transfectants requires a dominant selectable marker. A variety of such markers has been identified and is currently in use. However, many of these are not suitable for all cell types or require uniqu...

journal_title：BioTechniques

authors： Baccam M,Huberman E

更新日期：2003-06-01 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：:We describe a procedure for delivery of purified proteins into a variety of tissue culture cells using a new polycationic lipid preparation, LipofectAMINE. Several different proteins, with diverse physical properties, can be delivered into cells by this method. Compared with commercially available monocationic lipids,...

journal_title：BioTechniques

authors： Sells MA,Li J,Chernoff J

更新日期：1995-07-01 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：:Genome sequencing projects are either based on whole genome shotgun (WGS) or on a BAC-by-BAC strategy. Although WGS is in most cases the preferred choice, sometimes the BAC-by-BAC approach may be better because it requires a much simpler assembly process. Furthermore, when the study is limited to specific regions of t...

journal_title：BioTechniques

authors： Todesco S,Campagna D,Levorin F,D'Angelo M,Schiavon R,Valle G,Vezzi A

更新日期：2008-01-01 00:00:00

abstract：:To measure mutation load or to detect minimal residual disease, a robust method for identifying one mutant allele in the range of 10(6)-10(9) wild-type alleles would be advantageous. Herein, we present evidence that pyrophosphorolysis-activated polymerization (PAP) has the potential to provide a highly specific and ro...

journal_title：BioTechniques

authors： Liu Q,Sommer SS

更新日期：2000-11-01 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

authors： Bartel P,Chien CT,Sternglanz R,Fields S

更新日期：1993-06-01 00:00:00

abstract：:We have developed a rapid, sensitive, and quantitative 96-well microplate-based nonradioactive immunoprecipitation/kinase assay to evaluate mitogen-activated protein kinase (MAPK) activity. Three quantitative nonradioactive imunoprecipitation/kinase assays of MAPK were demonstrated on a 96-well microplate coated with ...

journal_title：BioTechniques

authors： Zhang L,Uder S,Juehne T,Brizzard B,Song K

更新日期：2002-02-01 00:00:00

abstract：:Several methods and computer programs have been developed for estimating the size of DNA fragments from gel electrophoresis. However, methods are lacking that may facilitate in optimization of gel conditions. In this article, a computer program called DNAOPT is described, which was developed to assist researchers in t...

journal_title：BioTechniques

authors： Raghava GP

更新日期：1995-02-01 00:00:00

abstract：:The Sequence Analysis Workshop (SAW) is an interactive program for sequence analysis of immunoglobulin variable domains. Sequences for SAW can be obtained from GenBank or from a standard text file. SAW can compare a variable domain to as many as 100 different sequences, calculate the extent of homology, sort the seque...

journal_title：BioTechniques

authors： Elgavish RA,Schroeder HW Jr

更新日期：1993-12-01 00:00:00

abstract：:Directed evolution is poised to change small molecule discovery and provide greater access to "drug space." Sarah Webb looks into the evolving drug discovery landscape. ...

journal_title：BioTechniques

authors： Webb S Ph D

更新日期：2017-12-01 00:00:00

abstract：:We have developed an improved subtractive hybridization method that provides a fast, simple and reliable isolation of desired different sequences from two compared DNA libraries, one of which contains all unwanted homologues (subtracter) and another contains certain desired heterologues (tester). The DNA library can b...

journal_title：BioTechniques

authors： Ying SY,Lin S

更新日期：1999-05-01 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：:Presepsin is a 13-kDa N-terminal glycoprotein of CD14. Previously, agitation-induced increases in presepsin levels have been reported; however, the mechanism remains poorly understood. In this study, we aimed to reveal the mechanism of presepsin increase. The agitated plasma or serum was separated using gel exclusion ...

journal_title：BioTechniques

authors： Morino G,Takahashi G,Kan S,Inoue Y,Sato K,Shirakawa K

更新日期：2021-01-29 00:00:00

abstract：:We describe a novel assay format for the Gal4-based yeast two-hybrid-system, in which the readout from three different reporter genes is measured sequentially in a single microplate. Activation of the URA3, MEL1, and lacZ reporters in response to a protein-protein interaction is monitored by measuring sequentially: (i...

journal_title：BioTechniques

authors： Evans DR,Swirsding KA,Taillon BE,Simons JF

更新日期：2004-11-01 00:00:00

abstract：:Thin spun-coat films (~4 nm thick) of graphene oxide (GO) constitute a versatile surface chemistry compatible with a broad range of technologically important sensor materials. Countless publications are dedicated to the nuances of surface chemistries that have been developed for sensors, with almost every material hav...

journal_title：BioTechniques

authors： Mulvaney SP,Stine R,Long NC,Tamanaha CR,Sheehan PE

更新日期：2014-07-01 00:00:00

abstract：:A modification of PCR-mediated gene synthesis strategy is introduced. This modification enables the synthesis of a gene from oligonucleotides comprising only one of the two strands. Bridging oligonucleotides (approximately 20-mers in length) complementary to the junctions of template strand oligonucleotides and two ou...

journal_title：BioTechniques

authors： Jayaraman K,Puccini CJ

更新日期：1992-03-01 00:00:00

abstract：:Identification of antimitotic molecules that affect tubulin dynamics is a multistep procedure. It includes in vitro tubulin polymerization assay, studies of a cell cycle effect, and general cytotoxicity assessment. To simplify this lengthy screening protocol, we have introduced and validated an assay system based on t...

journal_title：BioTechniques

authors： Semenova MN,Kiselyov A,Semenov VV

更新日期：2006-06-01 00:00:00

abstract：:A simple spectrophotometric method for measuring DNA in proliferating cells is described. The method is an adaptation of the widely used diphenylamine (DPA) reaction to examine DNA in cells grown in a 96-well plate. This assay was capable of detecting as little as 10 ng DNA and could be used to measure DNA in stored a...

journal_title：BioTechniques

authors： Natarajan N,Shambaugh GE 3rd,Elseth KM,Haines GK,Radosevich JA

更新日期：1994-07-01 00:00:00

abstract：:MicroRNAs (miRNAs) are ~22-nucleotide-long small non-coding RNAs that regulate the expression of protein-coding genes by base pairing to partially complementary target sites, preferentially located in the 3´ untranslated region (UTR) of target mRNAs. The expression and function of miRNAs have been extensively studied ...

journal_title：BioTechniques

authors： Persson H,Søkilde R,Pirona AC,Rovira C

更新日期：2017-08-01 00:00:00

abstract：:Gene expression profiles may offer more or additional information than classic morphologic- and histologic-based tumor classification systems. Because the number of tissue samples examined is usually much smaller than the number of genes examined, efficient data reduction and analysis methods are critical. In this rep...

journal_title：BioTechniques

authors： Xiong M,Jin L,Li W,Boerwinkle E

更新日期：2000-12-01 00:00:00

abstract：:A critical issue in transfection or co-transfection experiments is to define the appropriate controls. In most cases, a corresponding empty vector is used as one control. We report a paradoxical effect of empty mammalian expression vectors on different reporters. We have found that different empty vectors can inhibit ...

journal_title：BioTechniques

authors： Hu Q,Suzuki K,Hirschler-Laszkiewicz I,Rothblum LI

更新日期：2002-07-01 00:00:00

abstract：:In situ hybridization techniques typically employ chromogenic staining by enzymatic amplification to detect domains of gene expression. We demonstrate the previously unreported near infrared (NIR) fluorescence of the dark purple stain formed from the commonly used chromogens, nitro blue tetrazolium (NBT) and 5-bromo-4...

journal_title：BioTechniques

authors： Trinh le A,McCutchen MD,Bonner-Fraser M,Fraser SE,Bumm LA,McCauley DW

更新日期：2007-06-01 00:00:00

abstract：:A PCR-based method is described for the production of cDNA libraries and total cDNA probes from a few milligrams of tissue. Using a model system, we show how a PCR library and PCR probes can be used to identify genes expressed at different levels in two tissues. Small amounts of tissue derived from two plants, one inf...

journal_title：BioTechniques

authors： Bertiol DJ,Smoker M,Brown AC,Jones MG,Burrows PR

更新日期：1994-06-01 00:00:00

abstract：:Thymidine kinase 1 (TK1) is an enzyme involved in DNA precursor synthesis that has been used as a biomarker for prognosis and monitoring of different malignancies. In this study, we compared two immunoassays for measuring TK1 protein concentrations: the TK 210 ELISA (AroCell AB) and TK1 ELISA from Abcam. Overall, the ...

journal_title：BioTechniques

authors： Kumar JK,Holmgren S,Levedahl KH,Höglund M,Venge P,Eriksson S

更新日期：2020-06-01 00:00:00

abstract：:The colony formation assay (CFA) is the gold standard for measuring the effects of cytotoxic agents on cancer cells in vitro; however, in its traditional 6-well format, it is a time-consuming assay, particularly when evaluating combination therapies. In the interest of increased efficiency, the 6-well CFA was converte...

journal_title：BioTechniques

authors： Katz D,Ito E,Lau KS,Mocanu JD,Bastianutto C,Schimmer AD,Liu FF

更新日期：2008-02-01 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

authors： Major JG Jr

更新日期：1992-01-01 00:00:00

abstract：:Large genes present particular cloning difficulties, especially when expressed at relatively low levels. We describe a novel method, termed 3' rapid amplification of cDNA ends (RACE) walking, for the rapid determination of unknown 3' flanking sequence of a large cDNA. The technique is a derivative of the anchored PCR ...

journal_title：BioTechniques

authors： Park DJ,Pask AJ,Renfree MB,Graves JA

更新日期：2003-04-01 00:00:00