免疫学技术

相关专题 一般流程 甲醛处理细胞---收集细胞，超声破碎---加入目的蛋白的抗体，与靶蛋白-DNA复合物相互结合---加入ProteinA，结合抗体-靶蛋白-DNA复合物，并沉淀---对沉淀下来的复合物进行清洗，除去一些非特异性结合---洗脱，得到富集的靶蛋白-DNA复合物---解交联，纯化富集的DNA-片断---qPCR分析。在qPCR分析这一块，比较传统的做法是半定量-PCR。但是现在随着荧光定量PCR的普及，广州赛诚生物科技有限公司建议 ...

相关专题 HuH7细胞转入LXRα/RXRα表达质粒，并以抗LXRα抗体处理。经chip后对产物进行PCR分析。从实验结果可以明确LXRE2为与调控蛋白特异结合的位点。 标签: 广州赛诚生物 chip 转录调控 作者：广州赛诚生物 点击：次

一、实验步骤1、悬浮细胞样品把细胞及培养液一起收集到15 ml或者50 ml离心管中，1 500 rpm离心3 min，弃去上清；加入10 ml 冷PBS轻轻吹打，1 500 rpm，4℃离心3 min，弃去上清；反复2次；第二次弃去上清后，用1 ml 冷PBS重悬细胞，移入1.5 ml微型离心管，2 500 rpm（500 g），4℃离心5分钟，尽量去除所有残留上清，于-80度或者液氮冻存待用。在避免冻融的情况下，一般保存1年，建议尽快检测。2、贴壁细胞样品2.1 胰酶消化用胰酶将细胞消化 ...

Molecular hybridization is a useful technique for identifying specific target sequences even when they are present as a single copy in a complex population. It can be performed either on a solid matrix on which pure DNA (or RNA) is bound (blot hybridization) or on tissue sections (in situ hybridiza ...

With the advent of nonradioactive labels, in situ hybridization (ISH) has become a useful technique for the detection of viral DNA in infected tissue (1), mRNA expression (2), sex determination (3), human gene mapping (4), and interphase cytogenetics (5–8). For chromosomal mapping of genes by ISH, ...

The term in situ hybridization describes a wide range of techniques concerned with the detection of DNA or RNA sequences within individual cells, tissues, or on identifiable regions of chromosomes. The technique utilizes an ability to label DNA or RNA probes so that, following hybridization ...

The detection and quantitation of DNA adducts plays a central role in the determination of dose-response relationships for chemical carcinogens, mutagens, and chemotherapeutic agents in experimental laboratory investigations. Furthermore, it serves in molecular dosime ...

Monoclonal antibodies that are specific for sites of base modification in DNA have a number of applications in, for example, studies of carcinogenesis, drug action, or DNA repair (1). The production of antibodies of the appropriate specificity entails practical considerations that are s ...

Cytotoxic antibody-toxin conjugates made using antibodies and ribs some-inactivating proteins (RIPs) are prepared using chemical crosslinking methods (1,2 and this vol., Chapter 31). Gel permeation chromatography is used as a first step to purify conjugate molecules from the reac ...

Conjugates of antibodies with plant toxins, such as ricin and abrin, are potent cytotoxic agents that selectively eliminate target cells from mixed cell cultures in vitro, and have great promise as antitumor agents in cancer therapy (1). Ricin and abrin are protein toxins consisting of two dif ...

Twin-site ELISA is a simple technique for quantitation of specific proteins in cell or tissue extracts. The application of this method to fos and myc oncoproteins is described. There are two basic procedures: 1. Extraction of fos and myc proteins from biological material in a form suitable for imm ...

The great analytical power of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) makes it one of the most effective tools of protein chemistry and molecular biology. In the past, there have been many attempts to convert the technique from analytical to preparative s ...

The screening of a λgt11 library with monoclonal antibodies described here is a relatively uncomplicated procedure, but it requires a little introduction nevertheless. For simplicity, it is assumed that you have in your possession a library that is ready to screen, i.e., either a library boug ...

Whereas the expression of foreign genes in mammalian cells usually proves successful, the purification of gene products is often a difficult and time-consuming process. The availability of monoclonal antibodies to the foreign protein can greatly assist in small scale purification, ...

Several immunological procedures can be successfully carried out using nonpurified antibodies, such as unfractionated antisera, or ascitic fluid/culture supernatant containing monoclonal antibodies (MAbs). However, a much “cleaner” result can often be obtained if some fo ...

Morphologically, the lung is a complex organ containing over 40 different cell types (1). Although species and strain dependent, in the Fischer rat, the most common cell types include: endothelial, 33%; Type I, 6.5%; Type II, 12%; macrophages, 8%; ciliated and nonciliated bronchiolar epithelial ( ...

For flow cytometry, a suspension of single cells, free of large clumps and excess debris, is essential. The quality of the data obtained depends as much on the quality of the preparation as on that of the flow cytometer.

The isolation and subsequent study of hepatocytes in in vitro conditions was first transformed by Berry and Friend (1), who developed a collagenase liver perfusion method, allowing the isolation of large numbers of cells with high viability.

Immunomagnetic beads are uniform, polymer particles coated with a polystyrene shell that provides both a smooth hydrophobic surface to facilitate physical absorption of molecules, such as antibodies, and surface hydroxyl groups that allow covalent chemical binding of other bio ...

The enzyme immunoassay technique described below is a simple and sensitive method to detect left-handed Z-DNA formation in synthetic polynucleotides, recombinant plasmids, and native DNAs. This method utilizes the differences in the antigenic properties of right-handed B-DNA a ...