共查询到19条相似文献，搜索用时 250 毫秒
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采用重氮化法,将半抗原副品红(Pararosaniline,PA)分别与载体蛋白牛血清白蛋白(BSA)、人血清白蛋白(HSA)和卵清蛋白(OVA)偶联,反应得到免疫抗原和包被抗原,其偶联最佳结合比分别为8.8∶1,12.7∶1,6.9∶1,经紫外扫描光谱和蛋白电泳鉴定得到全抗原。用制备的抗原乳化后免疫小鼠获得了效价较高的多克隆抗体。为进一步制备抗孔雀石绿单克隆抗体打下基础。 相似文献
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建立多西环素人工抗原(Doxycycline,DOX)的合成及鉴定方法。采用戊二醛偶联法将DOX分别与载体牛血清白蛋白(BSA)及卵清蛋白(OVA)偶联制备免疫抗原(DOX-BSA)和包被抗原(DOX-OVA)。经SDS-PAGE电泳法、紫外可见吸收光谱鉴定,证实人工抗原成功合成,偶联比分别为8.6∶1和3.8∶1。结果表明建立了一种新的DOX抗原合成方法,为进一步制备特异性DOX抗体和建立检测食品中DOX的酶联免疫方法奠定了基础。 相似文献
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为了建立吲哚美辛(indomethacin,IDM)残留的免疫学检测方法,采用碳二亚胺法将载体蛋白牛血清白蛋白(BSA)和鸡卵清蛋白(OVA)分别与IDM偶联,人工合成免疫原(IDM-BSA)和包被原(IDM-OVA)。通过紫外分光光度法、SDS-PAGE、ELISA等方法鉴定人工抗原的合成效果;将IDM-BSA免疫小鼠,利用iELISA和icELISA来测定抗体血清效价。结果显示:IDM能与BSA和OVA成功偶联,通过免疫小鼠获得高效价的多克隆抗体,效价可达1∶6.4×10~4,半数抑制率为27.23 ng/mL。本研究结果为建立动物源性食品中IDM残留的免疫学检测方法奠定基础。 相似文献
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Six structural proteins of bovine parainfluenza-3 virus (PI-3V) labeled with [35S]-methionine could be resolved by polyacrylamide gel electrophoresis (PAGE). Five structural proteins of this virus had been previously reported. The 6 proteins found in this study were: L, a 180,000 (180 kD) molecular weight (MW) large protein; P, 83 kD phosphoprotein; HN, 69 kD hemagglutinin-neuraminidase glycoprotein; NP, 66 kD nucleocapsid protein; F, 55 kD fusion glycoprotein; and M, 38 kD matrix protein. Selective labeling with [2-3H]-mannose revealed only HN and F glycoprotein bands. A cellular actin protein (43 kD), associated with many enveloped viruses, was also found as a seventh protein in bovine PI-3V. 相似文献
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OBJECTIVE: To compare the electrophoretic pattern of serum proteins in clinically healthy adult camels (between 3 and 8 years of age) and camel calves (less than 3 months of age). DESIGN: Laboratory analysis of serum from healthy camels. PROCEDURE: Blood was collected from 30 healthy adult camels and 30 camel calves and the serum separated. Total protein of each serum sample was estimated by automated chemistry analyser. The proteins were fractionated by automated electrophoresis on agarose gel. RESULTS: Serum proteins migrated on the agarose gel as one albumin, two alpha (alpha1 and alpha2-globulins), two beta (beta1 and beta2-globulins) and one gamma-globulin fractions. In adult camels the mean concentration of total protein, albumin alpha1, alpha2, beta1, beta2 and gamma-globulins was 56.8 +/- 1.5, 30.7 +/- 0.8, 2.4 +/- 0.1, 3.2 +/- 0.1, 9.7 +/- 0.3, 3.4 +/- 0.2 and 8.6 +/- 0.3 g/L, respectively. These values in calves were 49.7 +/- 1.8, 23.7 +/- 0.8, 3.2 +/- 0.2, 3.1 +/- 0.2, 14.2 +/- 0.2, 4.0 +/- 0.2 and 4.1 +/- 0.2 g/L, respectively. CONCLUSION: The concentration of total proteins, albumin and gamma-globulins was higher (P < 0.05) in the adult camels than in camel calves. The concentrations of beta1 globulins was higher (P < 0.05) in calves as compared to adult camels. 相似文献
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Schellenberg S Mettler M Gentilini F Portmann R Glaus TM Reusch CE 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2008,22(2):273-281
BACKGROUND: Hypertension and proteinuria are commonly recognized in dogs with spontaneous hypercortisolism. There is, however, little information regarding the effect of exogenous glucocorticoids on blood pressure (BP) and proteinuria and whether these changes are reversible. HYPOTHESIS: Hydrocortisone administration increases systemic BP and urinary protein excretion, and these effects are reversible after hydrocortisone withdrawal. Animals: Six control dogs and 6 dogs treated with hydrocortisone. METHODS: BP, urine protein : creatinine ratio (UPC), microalbuminuria (MALB), urine albumin : creatinine ratio (UAC), and urine gel electrophoresis were evaluated before, during, and after administration of hydrocortisone (8 mg/kg PO q12h for 12 weeks) or placebo. RESULTS: BP and UPC increased substantially during hydrocortisone administration from 123 mmHg (range 114-136 mmHg) and 0.17 (0.15-0.28) to a maximum of 143 mmHg (128-148 mmHg) and 0.38 (0.18-1.78), respectively, on day 28. MALB developed in 4 dogs and UAC significantly increased in all dogs during hydrocortisone administration with the maximum on day 84. Both increases in BP and proteinuria were reversible and completely resolved within 1 month after stopping hydrocortisone administration. SDS-AGE revealed the proteinuria to be primarily albuminuria with a pronounced increase during hydrocortisone treatment. Furthermore, a protein of 25-30 kDa was found in male dogs, identified by mass spectrometry to be arginine esterase, the major secretory prostatic protein. CONCLUSIONS AND CLINICAL IMPORTANCE: Long-term hydrocortisone treatment results in significant but only mild increases in systemic BP and urinary protein excretion, which are both reversible within 1 month after discontinuation of hydrocortisone. 相似文献
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Kendal Harr John Harvey Robert Bonde David Murphy Mark Lowe Maya Menchaca Elsa Haubold Ruth Francis-Floyd 《Journal of zoo and wildlife medicine》2006,37(2):151-159
Manatees (Trichechus manatus latirostris) are afflicted with inflammatory and infectious disease secondary to human interaction, such as boat strike and entanglement, as well as "cold stress syndrome" and pneumonia. White-blood-cell count and fever, primary indicators of systemic inflammation in most species, are insensitive in diagnosing inflammatory disease in manatees. Acute phase-response proteins, such as haptoglobin and serum amyloid A, have proven to be sensitive measures of inflammation/infection in domestic large animal species. This study assessed diagnosis of generalized inflammatory disease by different methods including total white-blood-cell count, albumin: globulin ratio, gel electrophoresis analysis, C-reactive protein, alpha, acid glycoprotein, haptoglobin, fibrinogen, and serum amyloid A. Samples were collected from 71 apparently healthy and 27 diseased animals during diagnostic medical examination. Serum amyloid A, measured by ELISA, followed by albumin:globulin ratio, measured by plasma gel electrophoresis, were most sensitive in diagnosing inflammatory disease, with diagnostic sensitivity and specificity of approximately 90%. The reference interval for serum amyloid A is <10-50 microg/ml with an equivocal interval of 51-70 microg/ml. The reference interval for albumin:globulin ratio by plasma gel electrophoresis is 0.7-1.1. Albumin: globulin ratio, calculated using biochemical techniques, was not accurate due to overestimation of albumin by bromcresol green dye-binding methodology. Albumin:globulin ratio, measured by serum gel electrophoresis, has a low sensitivity of 15% due to the lack of fibrinogen in the sample. Haptoglobin, measured by hemoglobin titration, had a reference interval of 0.4-2.4 mg/ml, a diagnostic sensitivity of 60%, and a diagnostic specificity of 93%. The haptoglobin assay is significantly affected by hemolysis. Fibrinogen, measured by heat precipitation, has a reference interval of 100-400 mg/dl, a diagnostic sensitivity of 40%, and a diagnostic specificity of 95%. 相似文献
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Crivellente F Bonato M Cristofori P 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2008,37(1):73-78
BACKGROUND: Serum protein analysis in both humans and experimental animal species has so far been carried out by labor-intensive techniques, such as agarose gel electrophoresis (AGE). OBJECTIVE: The objective of this study was to evaluate capillary electrophoresis (CE) as an alternative technique to AGE for the analysis of serum proteins from healthy animals. METHODS: Blood samples were collected into tubes without anticoagulant from 6 fasted healthy male mice, rats, dogs, marmosets, and humans. Serum proteins were separated by CE using a technique standardized for the analysis of human proteins, and the results (efficiency, resolution, and precision) were compared with those obtained through AGE. RESULTS: Compared with AGE, CE resulted in narrower peaks and more peaks. The efficiency of protein separation by CE was significantly higher for all species, and resolution (R) was significantly higher in samples from dogs. Using rat serum, intraday reproducibility was lower for all protein fractions, and interday reproducibility was lower for most peaks, compared with AGE. CONCLUSIONS: We conclude that CE is a viable alternative to AGE for the determination of protein electrophoresis in a routine veterinary clinical pathology laboratory. The minimal sample requirement (2 microL), complete automation, and quantitative results make CE an especially valuable technique for protein analysis in experimental animal models. 相似文献
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Fayos M Couto CG Iazbik MC Wellman ML 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2005,34(4):397-400
BACKGROUND: Retired racing Greyhounds are becoming common as pets. Because of their unique physiology, results of routine laboratory tests are frequently outside the reference interval for dogs. Compared with other breeds, Greyhounds have low serum protein concentrations, but the concentrations of different serum protein fractions have not been reported. OBJECTIVES: Our objectives were to evaluate the results of serum protein electrophoresis (SPE) in healthy, retired racing Greyhounds and compare them with a control group of age- and gender-matched non-Greyhound dogs. METHODS: Agarose gel electrophoresis was done using a standard method; the gels were stained with amido black and scanned with a Cliniscan 2 densitometer (Helena Laboratories, Beaumont, TX, USA). Protein fractions were identified by visual inspection of the electrophoretogram. A Student's t-test assuming equal variances was used to compare the concentration of the different fractions between groups. RESULTS: The concentrations of total protein, total globulins, and alpha-1-, alpha-2-, beta-1-, and beta-2-globulins were significantly lower and the albumin to globulin (A:G) ratio was significantly higher in Greyhounds than in non-Greyhound dogs (P < .05). There was no significant difference in albumin or gamma-globulin concentrations. CONCLUSIONS: Low serum protein concentrations in Greyhounds are the result of low concentrations of a- and b-globulins. These results should be kept in mind when evaluating both healthy and sick Greyhounds. Additional studies are needed to identify the individual proteins associated with low alpha- and beta-globulin concentrations in Greyhounds. 相似文献
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Barbara Riond Bettina Wenger‐Riggenbach Regina Hofmann‐Lehmann Hans Lutz 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2009,38(1):73-77
Background: Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and α‐, β‐, and γ‐globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported. Objectives: The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm‐blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined. Methods: Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi‐automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within‐run and within‐assay precision. Data from warm‐blooded and draught horses were compared using the Mann–Whitney U test. Results: Within‐run and within‐assay CVs were <5% for all protein fractions. No significant difference was found between warm‐blooded and heavy draught horses and so combined reference intervals (2.5–97.5%) were calculated for total protein (51.0–72.0 g/L), albumin (29.6–38.5 g/L), α1‐globulin (1.9–3.1 g/L), α2‐globulin (5.3–8.7 g/L), β1‐globulin (2.8–7.3g/L), β2‐globulin (2.2–6.0 g/L), and γ‐globulin (5.8–12.7 g/L) concentrations, and albumin/globulin ratio (0.93–1.65). Conclusion: Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions. 相似文献
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采用聚丙烯酰胺凝胶电泳法对蓝马鸡和藏马鸡的血红蛋白、红细胞蛋白质、血清前白蛋白、白蛋白和运铁蛋白的蛋白谱进行了研究。结果发现 ,蓝马鸡和藏马鸡的血红蛋白 (HB)为由A1 和A2 2条区带组成的A1 A2 型 ;红细胞蛋白质 (EP)、血清白蛋白 (ALB)和运铁蛋白 (TF)分别呈单一的AA ,CC和FS。血清前白蛋白有Pr 1和Pr 2这 2种蛋白质组成 相似文献
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蛋白质组学以蛋白质组为研究对象,从整体上对生命载体进行研究,已成为后基因组时代的研究热点。目前,蛋白质组学技术主要包括双向凝胶电泳、生物质谱及生物信息学。双向凝胶电泳根据蛋白质的等电点和分子质量分离蛋白质,而质谱技术已成为鉴定蛋白质的极为灵敏而迅速的工具。由此得到的肽质量图谱结合准确全面的数据库技术,就使得新的蛋白质或多肽得以鉴定。近年来,蛋白质组研究技术已应用到多种生命科学领域中,在兽医学研究领域中尤其是兽药开发方面也将会有很广阔的应用前景。 相似文献