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991.
We describe a new immunoassay which combines an immunosorbent assay, Immunogold silver staining and epipolarization microscopy. Our new assay procedure features multiple samples on a single microscope slide, and high sensitivity of epipolarization microscope for detection of silver-enhanced colloidal gold as a final immunoassay product. We call the new immunoassay “on slide immunogold assay” (OSIGA). This new method uses biotinylated antibody and streptavidin-gold reaction with silver enhancement technique. With OSIGA it is possible to investigate 30 samples on a single microscopic slide. Our preliminary studies used 10-20 μ1 samples and detected nanogram quantities of a standardized protein solution. Unlike enzyme linked immunosorbent assay (ELISA), which has a limited time for reading the final color products, the OSIGA specimens can be dried or resin mounted for longer storage and future reference. 相似文献
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994.
Ajoy Basak Alain Boudreault Andrew Chen Michel Chrtien Nabil G. Seidah Claude Lazure 《Journal of peptide science》1995,1(6):385-395
Antiserum against an N-terminal sequence of murine prohormone convertase-1 (mPC1) incorporating the sequence immediatley following the junction between the putative pro-region and the active enzyme was obtained. This was accomplished using the multiple antigenic peptide (MAP) approach whereupon an 8-branched polylysine core to which are grafted multiple copies of a 16 amino acid peptide representing the N-terminal sequence of mPC1 (positions 84–99) was synthesized by solid-phase Fmoc chemistry. The ensuing peptide was purified and fully characterized by RP-HPLC, 1H-NMR, amino acid composition, peptide sequencing and ion-spray mass spectrometry. The immunological properties of the resulting antibodies in detecting recombinant PC1 in both crude and purified preparations were compared with antibodies raised against a similar N-terminal segment of PC1 but using the conventioanl method of peptide–carrier protein conjugation and also developed against a C-terminal fusion protein of PC1. Our data indicate that the MAP antibody was as efficient as both the amino and carboxy-terminal antibodies in qualitative as well as quantitative analysis of PC1 encoded protein by radioimmunoassay. Following an identical approach, antibodies against other prohormone convertases like furin, PC5/6 and PACE4 were also developed and subsequently applied to a number of biochemical and immunological studies. In each case, the ease of preparation and high immunogenicity of the MAP approach were confirmed and reside in the simplicity and rapidity with which a potent and useful antiserum is obtained. 相似文献
995.
This study investigated the compatibility of arbitrarily primed polymerase chain reaction (AP-PCR) and ribotyping in the characterization of Actinobacillus actinomycetemcomitans , a major pathogen in the mixed anaerobic microflora of human periodontitis. AP-PCR was performed directly on lysed bacterial colonies using a random-sequence 10-base oligonucleotide primer. Ribotyping was carried out by using purified bacterial chromosomal DNA digested with BglI. DNA fragments were separated electrophoretically, blotted onto a nylon membrane and hybridized with the plasmid pKK3535 containing the rRNA operon of Escherichia coli. The two genetic methods were evaluated on isolates from single individuals and from family members. Twelve AP-PCR types and 47 ribotypes were distinguished among 76 A. actinomycetemcomitans isolates of different serotypes. AP-PCR typing and ribotyping gave compatible results in 18 of 20 comparisons. Although AP-PCR detected less genetic heterogeneity in A. actinomycetemcomitans than ribotyping, the rapid and relatively simple AP-PCR technique seems to be sufficiently discriminative to be used in large scale epidemiological studies which preclude the application of the more laborious ribotyping technique. 相似文献
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997.
根瘤菌资源数据库(RRDB)的建立 总被引:1,自引:1,他引:0
在总结我室多年对根瘤菌资源调查与分类研究的基础上,建立了根瘤菌资源数据库(RRDB)。此库依据根瘤菌研究特点而设计,包括:基本信息、采集信息、保藏信息、回接信息、参考文献、性状信息、菌株说明等七个子库。目前收录了来自国内21个省或地区及国外部分研究单位提供的;并经全面性状分析与分类研究的286个菌株的信息,每个菌株设有寄主来源、固氮酶活性、碳源、氮源的利用等321个数据项。该库具有数据维护、查询检索、数据统计、输出打印等功能,引入并连接了聚类分析软件包-MINTS系统,可以满足当前研究的需要。 相似文献
998.
马铃薯Y病毒组病毒高产量提取方法的建立 总被引:11,自引:0,他引:11
本文报道了高产量提取芜菁花叶病毒(TuMV)、莴苣花叶病毒(LMV)、芋花叶病毒(DMV)和大豆花叶病毒(SMV)的提取方法。本方法通过使用高盐浓度的磷酸盐缓冲液以及在缓冲液中加入氯化镁和脲,并用TritonX-100作为澄清剂,替代常规使用的氯仿和正丁醇,成功的提取到了大量病毒粒子,上述四种病毒提取的得率分别是TuMV为173.3mg/kg病叶,LMV为96mg/kg病叶,SMV为199.2mg/kg病叶,DMV为176.6mg/kg病叶。 相似文献
999.
一种分离培养和观察微生物的简易方法 总被引:1,自引:0,他引:1
本文介绍了一种简易的分离、培养和观察微生物的方法(双载片法),适用于好氧菌及耐氧性厌氧菌。试验表明,此法的优点是:适于连续地观察和研究微生物的个体生长,了解其生活史;能快速简便地测定微生物的死亡率;能有效地分离纯种;为耐氧性厌氧菌的研究提供了一种便利的方法。 相似文献
1000.