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1.
Antisera were raised against tryptophan, 5-hydroxytryptophan, 5-hydroxytryptamine, 5-methoxytryptophan, and 5-methoxytryptamine, by conjugating each molecule to bovine serum albumin and to human serum albumin via glutaraldehyde, in such a way as to preserve the original part. Antibody specificity was tested with the enzyme-linked immunosorbent assay method. The specificity of each anti-indolealkylamine-glutaraldehyde antibody was established with competition experiments by using an adsorbed immunogenic conjugate and indolealkylamines either free or conjugated with poly-L-lysine. The nonconjugated compounds were poorly recognized. In the same way, the nonreduced conjugates always appeared less immunoreactive than the reduced ones. Calculated from the specificity study of each antiserum, the cross-reactivity ratios were found to be smallest for the most immunoreactive conjugates. Thus, a specific immune response was defined for each compound belonging to the same metabolic pathway. 相似文献
2.
Anthony D. Smith 《Archives of microbiology》1982,133(2):118-121
An indirect fluorescent antibody technique was used as a method of rapidly assessing and identifying sulphate-reducing bacteria. Five specific antisera and one polyvalent serum were raised and tested against 44 strains of the genera Desulfovibrio and Desulfotomaculum along with 4 control organisms. Immunofluorescence was found to be mainly strain specific with the sulphate-reducing bacteria although weak fluorescence was seen both within and between recognised groups. A polyvalent antiserum was successfully used to detect sulphate-reducing bacteria. No interference from 4 control organisms was found. 相似文献
3.
目的 分离纯化黄鳝血清免疫球蛋白,制备其兔抗血清,并检测抗血清的特异性。方法 用Protein A亲和层析的方法纯化黄鳝血清免疫球蛋白,通过SDS-聚丙烯酰胺凝胶电泳检测蛋白的纯度,免疫大耳白兔制备抗血清,利用免疫双扩散检测抗血清的效价,通过western blotting检测抗血清的特异性。结果 纯化了黄鳝血清免疫球蛋白,免疫双扩散法测定兔抗黄鳝免疫球蛋白血清效价为1∶32,western blotting结果显示抗血清具有很好的特异性。结论 成功纯化了黄鳝免疫球蛋白,制备了兔抗黄鳝IgM抗血清,为建立黄鳝的血清学检测系统奠定了基础。 相似文献
4.
Payakapong Waraporn Tittabutr Panlada Boonkerd Nantakorn 《World journal of microbiology & biotechnology》2003,19(9):981-983
Strain-specific antisera were produced against six Bradyrhizobium japonicum strains using two immunization procedures. These specific antisera were used for detection of bradyrhizobia in preserved soybean nodules. Antisera specific for two of these strains were either conjugated with a fluorescent dye or used with a fluorescent secondary antibody for identification of bradyrhizobia in soybean nodules that were preserved in four different storage conditions. Results show that soybean nodules dried in the oven, stored under room temperature, or at –20 °C are as suitable as fresh nodules for strain identification using fluorescent antisera. 相似文献
5.
A sensitive and specific radioimmunoassay (RIA) for the oxidised form of methionine5-enkephalin (Met5-Enk), Met5-Enk sulphoxide (Met5-Enk-S), has been developed. Antisera were raised in rabbits against Met5-Enk coupled to carrier proteins with glutaraldehyde or carbodiimide. Displacement of (125I) Met5-Enk bound to antiserum by Met5-Enk was poor, but Met5-Enk-S displayed good displacement suggesting that the Met5-Enk immunogen was oxidised to Met5-Enk-S and that the antisera were formed against this compound. The sensitivity of the RIA for Met5-Enk-S was 0.02 pmole/tube using the most sensitive antiserum. The antisera showed negligible cross-reactivity with leucine5-enkephalin and with both native and oxidised endorphins. Cross-reactivity was between 15% and 28% with the fragment Met5-Enk (2–5) sulphoxide and between 9% and 25% with D-Ala2-Met5-Enk sulphoxide. The antisera showed<0.01% cross-reactivity with other Met5-Enk fragments and naturally occurring neuropeptides. Tissue extracts were oxidised with hydrogen peroxide prior to assay. Met5-Enk-S immunoreactivity (IMR) was detected in brain, pituitary gland, pancreas, and intestine extracts of the rat, chicken, toad and teleost, and in cerebral-suboesophageal ganglion extracts of the snail. All tissue extracts showed parallelism in serial dilution to synthetic mammalian Met5-Enk-S, suggesting possible immunological identity. The results indicate that spontaneous oxidation of Met5-Enk immunogen occurs such that antisera are produced against the sulphoxide analogue of Met5-Enk, and may account for the relative insensitivity of some published RIAs using Met5-Enk standard. Our findings demonstrate a wide phylogenetic and anatomical distribution of Met5-Enk IMR. 相似文献
6.
猪脑组织提取液经SephadexG-50分子筛层析,S-SepharoseFastFlow阳离子交换柱层析及两次HPLC分离得到一分子量为12000,等电点pI7.1的多肽,并测定了其氨基酸组成和N末端部分序列:N-phe-Lys-Gly-Phe-Pro-Asp-Asp/(Lys)-Lys/(Asp)-Asp-Tyr.给昆明小鼠脑室注射或尾静脉注射该肽均能抑制吗啡引起的镇痛作用,其作用随着注射剂量的增大而增强.用BALB/C小鼠制备了该肽的抗血清,脑室注射此抗血清能明显逆转昆明小鼠对吗啡的耐受.因为这种来自猪脑的具有抗阿片镇痛作用的肽有99个氨基酸,所以简称此肽为AOP-99a(anti-oPioidpeptide). 相似文献
7.
以柯斯质粒pHC79为载体构建致肾盂肾炎大肠杆菌代表株E.coli J96染色体基因文库,获得两个能够表达粘附特性的重组柯斯质粒.进一步用鸟枪法亚克隆到载体pACYC184,得到三个阳性重组体.其中pCT10/E.coli K-12 p678-54能够稳定表达P菌毛和MRHA,分子量为14.6kb.用该克隆子免疫新西兰白兔,抗血清用带有载体质粒的受体菌(pACYC184/E.coli K-12 P678-54)吸收后,经菌毛粗提物的SDS-聚丙烯酰胺凝胶电泳及Western blotting和血凝抑制试验检测,其特异性仅针对致肾盂肾炎大肠杆菌的粘附基因群,可用于临床菌株的鉴定. 相似文献
8.
Jn M. Einarsson Patrick Joyce Yvette W. Kunz 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1995,112(4):589-598
The eye-specific C4-lactate dehydrogenase (LDH) and the widely distributed B4-LDH isozymes from the fish Oreochromis mossambicus were purified to homogeneity using DEAE Sepharose ion-exchange chromatography and oxamate-linked sepharose affinity-chromatography. Kinetic analysis was performed on pure B4- and C4-LDH. The Michaelis-Menten constant (Km) for B4-LDH and pyruvate was 32.3μM, for B4-LDH and lactate 717 μM for C4-LDH and pyruvate 14.1 μM and for C4-LDH and lactate 1898 μM. The pure C4-isozyme was subjected to SDS-polyacrylamide gel electrophoresis and the Coomassie Brilliant Blue-stained band injected into a rat to produce antiserum. The antiserum proved to be C4-LDH monospecific, which will allow using it to localize the isozyme in the retina at a light and electron microscopical level. 相似文献
9.
《Journal of receptor and signal transduction research》2013,33(1-4):253-266
ABSTRACTIn the search for yet unknown subtypes of GABAB receptors, the subunit architecture of GABAB receptors in the retina was analyzed using selective antisera. Immunopurification of the splice variants GABAB1a and GABAB1b demonstrated that both were associated with GABAB2. Quantitative immunoprecipitation experiments indicated that practical the entire GABAB receptor population in the retina consists of the receptor subtypes GABAB1a/GABAB2 and GABAB1b/GABAB2, although low levels of GABAB1c/GABAB2 cannot be excluded. The data rule out the existence of GABAB receptors containing the splice variants GABAB1d and GABAB1e. Moreover, no evidence for homomeric GABAB1 receptors was found. Among the splice variants, GABAB1a is by far the predominant one in neonatal and adult retina, whereas GABAB1b is expressed only late in postnatal development and in the adult retina. Since GABAB1a is expressed at high levels before functional synapses are formed, this specific receptor subtype might be involved in the maturation of the retina. Finally, subcellular fractionation demonstrated that GABAB1a, but not GABAB1b, is present in postsynaptic densities, suggesting a differential pre- and postsynaptic localisation of both splice variants. 相似文献
10.