Abstract：
:Screening methods for chemotherapeutic agents usually rely on the cytotoxic properties of the drugs. However, agents that inhibit invasion may have more efficacy and cause fewer side effects. Various cellular invasion assays have been used to evaluate these types of compounds, including the modified Boyden chamber, monolayer wound models and Matrigel outgrowth assays. In this report, we have combined the use of the Matrigel outgrowth assay with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) visualization and cell viability dye to visualize invasive cells on Matrigel without magnification. Extraction of the dye's formazan byproduct allows cell viability to be assessed. Using several invasive and noninvasive cell lines, the utility of the method for various target cells was verified. Several established chemotherapeutic agents were also screened for their anti-invasive and/or cytotoxic effects when cultured on Matrigel. Our results suggest that this method may be an easy, inexpensive and nonradioactive alternative for both enumerating cells on Matrigel and screening various tumor cell lines treated with chemotherapeutic agent to look for compounds with noncytotoxic but anti-invasive properties.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Sasaki CY,Passaniti Adoi
10.2144/98246cr04subject
Has Abstractpub_date
1998-06-01 00:00:00pages
1038-43issue
6eissn
0736-6205issn
1940-9818journal_volume
24pub_type
杂志文章相关文献
BIOTECHNIQUES文献大全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：:We describe a general method for making template DNA for sequencing of PCR products. The procedure may be particularly useful for PCR products where minimal sequence information is known or as an alternative to primer walking when sequencing long PCR products. A cassette containing the hybridization site for the M13 s...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-11-01 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：:We describe a method for rapid radioactive labeling of PCR product. The method, employing the Klenow fragment of DNA polymerase I, consumes little product, requires no product purification and takes under 30 minutes. ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1991-08-01 00:00:00
abstract：:We describe a rapid and sensitive method for the detection of nucleotide sequence variation that can be used for large-scale screening of population markers. Denaturing gradient gel electrophoresis (DGGE) detects sequence variants of amplified fragments by the differences in their melting behavior. DGGE detects most s...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/99275rr02
更新日期：1999-11-01 00:00:00
abstract：:Recombinant alphaviruses have been used as vehicles for delivery and expression of heterologous genes in mammalian, avian and insect cell lines. We have used a Sindbis replicon virus (Sinreplac) able to express the E. coli lacZ gene to compare the efficiency of transduction in one insect, six mammalian cell lines and ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/96213rr02
更新日期：1996-09-01 00:00:00
abstract：:Sequencing of the human genome in combination with computational annotation has provided tremendous data on predicted genes. However, for most of them, no corresponding cDNAs are available yet. Furthermore, even where cDNA clones were obtained, gene transcripts often have many different splice variants that are not co...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/04371DD02
更新日期：2004-07-01 00:00:00
abstract：:To avoid the time-consuming reprobing process in hybridization analysis, signal-distinguishable probes (32P, 35S or antigenic hapten-labeled DNA) can be added to the same hybridization mixture. After hybridization, an unambiguous result can be obtained by differential or sequential autoradiography. ...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1994-04-01 00:00:00
abstract：:Loop-mediated isothermal amplification (LAMP) is a rapid and reliable sequence-specific isothermal nucleic acid amplification technique. To date, all reported real-time detection methods for LAMP have been restricted to single targets, limiting the utility of this technique. Here, we adapted standard LAMP primers to c...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/0000113902
更新日期：2012-08-01 00:00:00
abstract：:SOP3 is a web-based software tool for designing oligonucleotide primers for use in the analysis of single nucleotide polymorphisms (SNPs). Accessible via the Internet, the application is optimized for developing the PCR and sequencing primers that are necessary for Pyrosequencing. The application accepts as input gene...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/05381RR01
更新日期：2005-01-01 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
pub_type： 杂志文章
doi：10.2144/03352pt02
更新日期：2003-08-01 00:00:00
abstract：:A new method has been developed to purify and detritylate milligram amounts of synthetic oligonucleotides. Dimethoxytrityl oligonucleotides from 15 to 100 nucleotides in length are applied in triethylammonium acetate or concentrated ammonium hydroxide to a disposable chromatographic cartridge, the NENSORB PREP Nucleic...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1990-04-01 00:00:00
abstract：:The purpose of this research was to compare three bioreactor systems for the pilot-scale production of monoclonal antibodies (MAbs). We needed to produce gram quantities of murine MAbs against human prostatic acid phosphatase for use in fragmentation, radiolabeling and in vivo radio-imaging studies. The stable hybrido...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1993-10-01 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：: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
pub_type： 杂志文章
doi：10.2144/04374ST02
更新日期：2004-10-01 00:00:00
abstract：:We propose two methods for characterizing the spatio-temporal behavior of cell populations in culture. The first method, image auto-correlation microscopy (IACM), allows us to characterize the variation in the number of objects as a function of time, thus enabling the quantification of the clustering properties of cel...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000112478
更新日期：2007-07-01 00:00:00
abstract：:A magnetic bead-based system for DNA isolation utilizing monodisperse beads was tested with the aim of producing a general approach for PCR-ready DNA. This commercially available system was originally designed for isolating PCR-ready DNA from human whole blood. We tested diverse organisms belonging to the major groups...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/97223rr01
更新日期：1997-03-01 00:00:00
abstract：:This paper reports a novel method for the identification of nucleic acid target sequences when these targets have high sequence identity. Homologous genes are currently identified by sequencing. We hypothesize that by primer extension in the presence of selected nucleotides, genes with similar sequence can be identifi...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1993-06-01 00:00:00
abstract：:Extracellular vesicles (EVs) are spherical membrane structures released by most cells. These highly conserved mediators of intercellular communication carry proteins, lipids, and nucleic acids, and transfer these cellular components between cells by different mechanisms, such as endocytosis, macropinocytosis, or fusio...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114371
更新日期：2016-01-01 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：:We describe a bead-based, multiplexed, oligonucleotide ligation assay (OLA) performed on the Luminex flow cytometer. Differences between this method and those previously reported include the use of far fewer beads and the use of a universal oligonucleotide for signal detection. These innovations serve to significantly...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000112960
更新日期：2008-11-01 00:00:00
abstract：:Piezoelectric dispensing of proteins from borosilicate glass capillaries is a popular method of protein biochip fabrication that offers the advantages of sample recovery and noncontact with the printing substrate. However, little regard has been given to the quantitative aspects of dispensing minute volumes (1 nL or l...
journal_title：BioTechniques
pub_type：
doi：10.2144/03342mt02
更新日期：2003-02-01 00:00:00
abstract：:Next-generation sequencing (NGS) of multigene panels performed for genetic clinical diagnostics requires 100% coverage of all targeted genes. In the genetic diagnostics laboratory, coverage gaps are typically filled with Sanger sequencing after NGS data are collected and analyzed. Libraries prepared using the hybridiz...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114217
更新日期：2014-10-01 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
pub_type： 新闻
doi：10.2144/000114616
更新日期：2017-12-01 00:00:00
abstract：:Millions of museum specimens are integral to biodiversity studies; however, DNA degradation may limit the ability to obtain DNA sequences. In this study, a degradation analysis model for Lepidoptera specimens was established. Based on this model, we revealed the characteristics of DNA fragment distribution caused by e...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/btn-2019-0166
更新日期：2020-03-01 00:00:00
abstract：:We have demonstrated that efficient polymerase chain reaction amplifications from chromosomal DNA can be carried out using whole bacterial cells as the starting material. Cells from the liquid or solid cultures can be used directly, without any pre-treatment, thus eliminating the need for DNA isolation. ...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1991-01-01 00:00:00
abstract：:Few studies have focused on the significance of ras protein levels in human malignancy, in part because of the inherent difficulty in quantitation of the ras gene product. We have developed a method for the enzymatic determination of the ras gene product and have used this method for the quantitation of ras gene produ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1990-08-01 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
abstract：:Microarray technologies have made possible comprehensive analyses of nucleic acid sequence and expression. However, the technology to obtain efficiently high-quality RNA and DNA suitable for array analysis from purified populations of neoplastic cells from human tissues has not been well addressed. Microdissection can...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/02324rr06
更新日期：2002-04-01 00:00:00
abstract：:Comprehensive analysis of DNA methylation patterns is critical for understanding the molecular basis of many human diseases. While hundreds of PCR-based DNA methylation studies are published every year, the selection and implementation of appropriate methods for these studies can be challenging for molecular genetics ...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：10.2144/000114087
更新日期：2013-10-01 00:00:00
