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121.
An improved Enzyme Immunoassay for Myelin Basic Protein is described. Myelin Basic Protein covalently attached to glass balls, and Myelin Basic Protein in samples compete with each other for binding of a peroxidase conjugated anti Myelin Basic Protein antibody. The peroxidase activity on the balls is then inversely proportional to the amount of Myelin Basic Protein in the sample. A detection limit of 0.6 ng/ml is demonstrated for diluent or spinal fluid. For plasma a dilution step increases this to 1.8 ng/ml. Both the coated balls and the peroxidase conjugate are stable for long periods. The assay requires no expensive equipment. Although the assay appears to be valid for subcellular fractions spinal fluid and plasma, successful detection of Myelin Basic Protection peptides in clinical samples may require careful selection of suitable antisera. The assay would be very suitable for eventual use with an appropriate monoclonal antibody. 相似文献
122.
The nonelectrophoretic transfer of proteins from thin (0.5 mm) isoelectric focusing gels to nitrocellulose was complete in 1 h. Blotting was bidirectional with 60% of the protein transferred to the top blot and the remaining 40% to the bottom blot. The use of nondenaturing transfer buffers permits proteins to be blotted in the native state. Immunological determinants are preserved and enzyme activity can be detected on the blots. 相似文献
123.
本文建立了组织型纤溶酶原激活剂(t-PA)活力的发光固相测定方法。用氨基乙基丁基异鲁米诺(ABEI)标记纤维蛋白原(Fg),在一定条件下,t-PA作用于固相(包被ABEI-Fg),产生纤维蛋白的降解产物。测定可溶性降解产物的发光强度,即能计算t-PA活性。该方法的标准曲线范围对t-PA为0.156IU/mL~40IU/mL。灵敏度可达0.156IU/mL。回收率为98.6%(n=27)。批内批间变异系数分别为6.6%及10.3%。该方法曾用于检测细胞培养液中提取t-PA样品及t-PA基因表达时培养液中t-PA的活性。也曾用于检测从组织中纯化t-PA样品及血浆中t-PA活性的测定。文中讨论了该方法与其它方法优缺点的比较。 相似文献
124.
B. M. Wagland W. O. Jones L. Hribar T. Bendixsen D. L. Emery 《International journal for parasitology》1992,22(8):1183-1185
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1992. A new simplified assay for larval migration inhibition. International Journal for Parasitology 22: 1183–1185. A simple method is described for the in vitro detection of substances that impair the motility of third-stage larvae of gastro-intestinal nematodes. The test is based on the ability of larvae to freely migrate through selected mesh sizes of nylon sieves and the reduced ability of larvae to migrate after preincubation with, and in the presence of, substances that inhibit or reduce larval motility. 相似文献
125.
Marc Mercken† Mark Vandermeeren† Ursula Lübke‡ Jan Six Jef Boons† Eugène Vanmechelen re Van De Voorde Jan Gheuens† 《Journal of neurochemistry》1992,58(2):548-553
Immunoaffinity chromatography with a monoclonal antibody produced against bovine tau protein was used to purify tau proteins from human brain. Fifty grams of brain tissue yielded approximately 2 mg of pure tau proteins. The affinity-purified human tau was used to produce a high-titered rabbit anti-human tau serum. The monoclonal anti-tau antibody and the polyclonal rabbit anti-tau serum were then used to construct a sandwich enzyme-linked immunosorbent assay for detection of human tau proteins, with a sensitivity of 1 ng/ml. 相似文献
126.
Measurement of bacterial random motility and chemotaxis coefficients: I. Stopped-flow diffusion chamber assay 总被引:4,自引:0,他引:4
Bacterial chemotaxis, the directed movement of a cell population in response to a chemical gradient, plays a critical role in the distribution and dynamic interaction of bacterial populations in nonmixed systems. Therefore, in order to make reliable predictions about the migratory behavior of bacteria within the environment, a quantitative characterization of the chemotactic response in terms of intrinsic cell properties is needed.The design of the stopped-flow diffusion chamber (SFDC) provides a well-characterized chemical gradient and reliable method for measuring bacterial migration behavior. During flow through the chamber, a step change in chemical concentration is imposed on a uniform suspension of bacteria. Once flow is stopped, diffusion causes a transient chemical gradient to develop, and bacteria respond by forming a band of high cell density which travels toward higher concentrations of the attractant. Changes in bacterial spatial distributions observed through light scattering are recorded on photomicrographs during a 10-min period. Computer-aided image analysis converts absorbance of the photographic negatives to a digital representation of bacterial density profiles. A mathematical model (part II) is used to quantitatively characterize these observations in terms of intrinsic cell parameters: a chemotactic sensitivity coefficient, mu(0), from the aggregate cell density accumulated in the band and a random motility coefficient, mu, from population dispersion in the absence of a chemical gradient.Using the SFDC assay and an individual-cell-based mathematical model, we successfully determined values for both of these population parameters for Escherichia coli K12 responding to fucose. The values obtained were mu = 1.1 +/- 0. 4 x 10(-5) cm(2)/s and chi(o) = 8 +/- 3 +/- 10(-5) cm(2)/s. We have demonstrated a method capable of determining these parameter values from the now validated mathematical model which will be useful for predicting bacterial migration in application systems. 相似文献
127.
128.
An immobilized cyclic multi-step enzyme system -- the urea cycle 总被引:1,自引:0,他引:1
129.
130.
Johan Stjernschantz Dale Gregerson Larry Bausher Marvin Sears 《Journal of neurochemistry》1982,38(5):1323-1328
A solid phase enzyme-linked immunosorbent assay for quantitation of substance P is presented. The assay measures the capacity of soluble substance P to compete with the solid phase antigen for a limited quantity of specific substance P antibody. The solid-phase antigen consists of a synthetic substance P.poly-D-glutamic acid conjugate coated to polystyrene micro-ELISA plate wells. Soluble substance P and antibodies to substance P are first preincubated together and then added to the wells containing solid-phase antigen. Subsequently the wells are incubated with anti-antibodies conjugated to alkaline phosphatase. The wells are finally incubated with p-nitrophenyl phosphate an the absorbance is read in a spectrophotometer 16--24 hr after the start of the assay. The threshold for detection of substance P was 5--10 pg per well (0.25 ml). Substance P was extracted from rabbit eyes and the values obtained with the present method are compared with previously reported values based on radioimmunoassay. 相似文献