scholarly journals ANTICORPOS POLICLONAIS PARA O IMUNODIAGNÓSTICO DE VÍRUS BOVINO

2019 ◽  
Vol 20 ◽  
Author(s):  
Tatiane Goulart de Lima ◽  
Luana Marchi Quadros ◽  
Leandro Abel Mallmann ◽  
Mário Celso Sperotto Brum
Keyword(s):  
Slot Blot

Resumo O objetivo do estudo foi produzir anticorpos policlonais para vírus bovino e testar a reatividade em imunoensaios como imunofluorescência, imunoperoxidase e slot blot. Como imunógeno utilizou-se cepas e/ou isolados dos herpesvírus bovino tipo 1, 2 e 5 (BoHV-1, BoHV-2 e BoHV-5), vírus da diarreia viral bovina (BVDV), vírus respiratório sincicial bovino (BRSV), vírus da língua azul (BTV) e vírus vaccina (VACV) amplificados em cultivo celular. Os coelhos foram imunizados em intervalos regulares e o soro hiperimune produzido foi utilizado como anticorpo primário nos imunoensaios. A diluição de trabalho para cada antissoro foi determinada por diluição seriada limitante e variaram entre 1:800 e 1:51.200. As maiores diluições foram observadas para imunoperoxidase, quando comparadas com a imunofluorescência e slot blot. Diluições menores que 1:800 foram associadas com aumento de reações inespecíficas. Os antissoros anti-BoHV-1, -BoHV-5, -BVDV e -BRSV reagiram frente a isolados heterólogos em ensaios de imunofluorescência e imunoperoxidase. Conclui-se que os antissoros produzidos possuem elevadas concentrações de anticorpos específicos para os vírus e é uma alternativa para a utilização em imunoensaios.

Using genomic slot blot hybridization to assess intergeneric Saccharum x Erianthus hybrids (Andropogoneae – Saccharinae)

Genome ◽  
1997 ◽  
Vol 40 (4) ◽  
pp. 428-432 ◽  
Author(s):  
P. Besse ◽  
C. L. McIntyre ◽  
D. M. Burner ◽  
C. G. de Almeida
Keyword(s):  
Genomic Dna ◽  
Blot Hybridization ◽  
Female Parent ◽  
Saccharum Officinarum ◽  
Rapid Screening ◽  
Hybridization Technique ◽  
Slot Blot ◽  
Saccharum Species ◽  
Slot Blot Hybridization

The use of genomic slot blot hybridization enabled the differentiation of hybrids from selfs in Saccharum × Erianthus intergeneric crosses in which Saccharum was used as the female parent. Based on the genomic in situ hybridization technique, slot blots of DNA from the parents and the progeny were blocked with the Saccharum parent DNA and hybridized with the labelled male Erianthus genomic DNA. This technique allowed a rapid screening for hybrids and was sensitive enough to detect a 1/20 dilution of Erianthus in Saccharum DNA, which should enable the detection of most partial hybrids. The genomic slot blot hybridization technique was shown to be potentially useful for assessing crosses involving Saccharum species with either Old World Erianthus section Ripidium or North American Erianthus (= Saccharum) species. The effectiveness of the technique was assessed on 144 progeny of a Saccharum officinarum × Erianthus arundinaceus cross, revealing that 43% of the progeny were selfs. The importance of this test as a tool to support intergeneric breeding programs is discussed.Key words: slot blot, Erianthus, genomic DNA, Saccharum, sugarcane.

scholarly journals A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin fromClostridium septicum

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3407 ◽  
Author(s):  
Lorena Vázquez-Iglesias ◽  
Borja Estefanell-Ucha ◽  
Leticia Barcia-Castro ◽  
María Páez de la Cadena ◽  
Paula Álvarez-Chaver ◽  
...  
Keyword(s):  
Protein Purification ◽  
Polyclonal Antibodies ◽  
Clinical Intervention ◽  
Farm Animals ◽  
Alpha Toxin ◽  
Slot Blot ◽  
Clostridium Septicum ◽  
Time Saving ◽  
Specific Antibodies

Clostridium septicumproduces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced byClostridium septicum.This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin ofClostridium septicumand produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot.

scholarly journals Using a CCD Camera Imaging System as a Recording Device to Quantify Human DNA by Slot Blot Hybridization

BioTechniques ◽  
2001 ◽  
Vol 30 (3) ◽  
pp. 680-685 ◽  
Author(s):  
Bruce Budowle ◽  
William R. Hudlow ◽  
Steven B. Lee ◽  
Leonard Klevan
Keyword(s):  
Imaging System ◽  
Blot Hybridization ◽  
Ccd Camera ◽  
Recording Device ◽  
Slot Blot ◽  
Camera Imaging ◽  
Human Dna ◽  
Slot Blot Hybridization

Slot blot immunoassay as a tool for plasmid-encoded toxin detection in enteroaggregative Escherichia coli culture supernatants

2006 ◽  
Vol 55 (2) ◽  
pp. 101-106 ◽  
Author(s):  
Andréa Bernardes Vilhena-Costa ◽  
Roxane Maria Fontes Piazza ◽  
Júlia Mitico Nara ◽  
Luiz Rachid Trabulsi ◽  
Marina Baquerizo Martinez
Keyword(s):  
Escherichia Coli ◽  
Slot Blot ◽  
Toxin Detection ◽  
Culture Supernatants

Absence of Changes in Metallothionein RNA in the Rat Testes Made Refractory to Cadmium Toxicity by Zinc Pretreatment

1994 ◽  
Vol 13 (1) ◽  
pp. 65-67 ◽  
Author(s):  
Zakaria Z. Wahba ◽  
Mark Steven Miller ◽  
Michael P. Waalkes
Keyword(s):  
Low Dose ◽  
Interstitial Cell ◽  
Cadmium Toxicity ◽  
Cdna Probe ◽  
Testicular Toxicity ◽  
Slot Blot ◽  
Tissue Resistance ◽  
Blot Technique ◽  
Induced Tolerance ◽  
Tolerance To Cadmium

Testicular toxicity and interstitial cell tumours induced by cadmium are prevented by zinc or by low dose cadmium pretreatments. The mechanism of this tolerance is unknown, though metallothionein (MT) is thought to play a role in tissue resistance to cadmium toxicity. Thus, the possible involvement of the testicular MT gene in metal-induced tolerance to cadmium toxicity was studied. Rats were pretreated with zinc (1.0 mmol kg-1, s.c.). Histological examination of the testes indicated such pretreatments prevented the necrotizing effects of subsequent doses of cadmium (20 ?mol kg-1, s.c.) administered 24 h later. RNA was extracted from testes or liver 24 h after zinc pretreatment, and analysed by the slot blot technique using the p2A10 cDNA probe to the MT gene. Zinc pretreatment had little effect on MT RNA in the testes, and such pretreatments did not alter testicular cadmium-binding protein capacity. In contrast, RNAs derived from livers of zinc pretreated rats showed marked increases in MT RNA and MT protein. Hence, the testicular MT gene does not appear to play a major role in the induced tolerance to cadmium toxicity and carcinogenesis generated by zinc.

Serum laminin assayed by Slot-Blot-ELISA in patients with combined viral hepatitis C and schistosomiasis

2006 ◽  
Vol 39 (6) ◽  
pp. 652-657 ◽  
Author(s):  
Samir El-Masry ◽  
Mahmoud Lotfy ◽  
Mohamed El-Shahat ◽  
Gamal Badra
Keyword(s):  
Hepatitis C ◽  
Viral Hepatitis ◽  
Slot Blot ◽  
Viral Hepatitis C
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