Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis

Level of superoxide dismutase (SOD) in the seedlings of downy mildew resistant pearl millet genotype (IP 18293) increased 3-fold upon inoculation with Sclerospora graminicola. Mn-SOD was purified from pearl millet by ion-exchange and gel filtration chromatography. Polyclonal antibody was raised in r...

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Autores principales:	Urs Babitha,Mysore Pandurangaraj, Madhu,Devaiah, Sripathi P,Harishchandra, Shekar Shetty,Hunthrike
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2006
Materias:	DIBA downy mildew ELISA immunofluorescence manganese superoxide dismutase pearl millet Sclerospora graminicola Western blot
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000200007
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spelling	oai:scielo:S0717-345820060002000072006-12-11Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosisUrs Babitha,Mysore PandurangarajMadhu,DevaiahSripathi P,HarishchandraShekar Shetty,Hunthrike DIBA downy mildew ELISA immunofluorescence manganese superoxide dismutase pearl millet Sclerospora graminicola Western blot Level of superoxide dismutase (SOD) in the seedlings of downy mildew resistant pearl millet genotype (IP 18293) increased 3-fold upon inoculation with Sclerospora graminicola. Mn-SOD was purified from pearl millet by ion-exchange and gel filtration chromatography. Polyclonal antibody was raised in rabbit against Mn-SOD of pearl millet. Anti-Mn SOD had an antibody titer of 1:20,000. ELISA results revealed a 1.5-fold increase in pure Mn-SOD preparation compared to the crude preparation. The antibody reactivity as detected by ELISA revealed that Mn-SOD is higher at 4 hrs post-inoculation in resistant pearl millet seedlings. The reactivity also showed the Mn-SOD is more in roots of inoculated resistant seedlings. SOD reactivity was also determined by DIBA (Dot-immunobinding assay). The results suggested that the Mn-SOD antibody reacted more strongly with crude SOD of the inoculated resistant seedlings than inoculated resistant seedlings. Western blot analysis revealed the induction of 35 kDa SOD protein in the resistant genotype. Its appearance on western blot coincided with high SOD activity. Immunolocalization experiments showed that SOD protein was abundant in the vascular bundles. The antibody produced to Mn-SOD was specific as judged by ELISA, DIBA, western blot and immunofluorescence assays. The work described here investigates the possibility of using the serological techniques to assess the reactivity of antibody with the SOD antigen.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.9 n.2 20062006-04-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000200007en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	DIBA downy mildew ELISA immunofluorescence manganese superoxide dismutase pearl millet Sclerospora graminicola Western blot
spellingShingle	DIBA downy mildew ELISA immunofluorescence manganese superoxide dismutase pearl millet Sclerospora graminicola Western blot Urs Babitha,Mysore Pandurangaraj Madhu,Devaiah Sripathi P,Harishchandra Shekar Shetty,Hunthrike Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis
description	Level of superoxide dismutase (SOD) in the seedlings of downy mildew resistant pearl millet genotype (IP 18293) increased 3-fold upon inoculation with Sclerospora graminicola. Mn-SOD was purified from pearl millet by ion-exchange and gel filtration chromatography. Polyclonal antibody was raised in rabbit against Mn-SOD of pearl millet. Anti-Mn SOD had an antibody titer of 1:20,000. ELISA results revealed a 1.5-fold increase in pure Mn-SOD preparation compared to the crude preparation. The antibody reactivity as detected by ELISA revealed that Mn-SOD is higher at 4 hrs post-inoculation in resistant pearl millet seedlings. The reactivity also showed the Mn-SOD is more in roots of inoculated resistant seedlings. SOD reactivity was also determined by DIBA (Dot-immunobinding assay). The results suggested that the Mn-SOD antibody reacted more strongly with crude SOD of the inoculated resistant seedlings than inoculated resistant seedlings. Western blot analysis revealed the induction of 35 kDa SOD protein in the resistant genotype. Its appearance on western blot coincided with high SOD activity. Immunolocalization experiments showed that SOD protein was abundant in the vascular bundles. The antibody produced to Mn-SOD was specific as judged by ELISA, DIBA, western blot and immunofluorescence assays. The work described here investigates the possibility of using the serological techniques to assess the reactivity of antibody with the SOD antigen.
author	Urs Babitha,Mysore Pandurangaraj Madhu,Devaiah Sripathi P,Harishchandra Shekar Shetty,Hunthrike
author_facet	Urs Babitha,Mysore Pandurangaraj Madhu,Devaiah Sripathi P,Harishchandra Shekar Shetty,Hunthrike
author_sort	Urs Babitha,Mysore Pandurangaraj
title	Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis
title_short	Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis
title_full	Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis
title_fullStr	Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis
title_full_unstemmed	Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis
title_sort	production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2006
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000200007
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Production of polyclonal antisera to manganese superoxide dismutase expressed in downy mildew resistant pearl millet and its application for immunodiagnosis

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