A strategy for screening monoclonal antibodies for Arabidopsis flowers

Qian Shi, Lian Zhou, Yingxiang Wang, Hong Ma

Research output: Contribution to journalArticle

Abstract

The flower is one of the most complex structures of angiosperms and is essential for sexual reproduction. Current studies using molecular genetic tools have made great advances in understanding flower development. Due to the lack of available antibodies, studies investigating the localization of proteins required for flower development have been restricted to use commercial antibodies against known antigens such as GFP, YFP, and FLAG. Thus, knowledge about cellular structures in the floral organs is limited due to the scarcity of antibodies that can label cellular components. To generate monoclonal antibodies that can facilitate molecular studies of the flower, we constructed a library of monoclonal antibodies against antigenic proteins from Arabidopsis inflorescences and identified 61 monoclonal antibodies. Twenty-four of these monoclonal antibodies displayed a unique band in a western blot assay in at least one of the examined tissues. Distinct cellular distribution patterns of epitopes were detected by these 24 antibodies by immunofluorescence microscopy in a flower section. Subsequently, a combination of immunoprecipitation and mass spectrometry analysis identified potential targets for three of these antibodies. These results provide evidence for the generation of an antibody library using the total plant proteins as antigens. Using this method, the present study identified 61 monoclonal antibodies and 24 of them were efficiently detecting epitopes in both western blot experiments and immunofluorescence microscopy. These antibodies can be applied as informative cellular markers to study the biological mechanisms underlying floral development in plants.

Original languageEnglish (US)
Article number270
JournalFrontiers in Plant Science
Volume8
DOIs
StatePublished - Feb 28 2017

Fingerprint

monoclonal antibodies
Arabidopsis
screening
flowers
antibodies
fluorescence microscopy
flowering
epitopes
Western blotting
antigens
cell structures
plant proteins
sexual reproduction
molecular genetics
Angiospermae
inflorescences
proteins
mass spectrometry
assays

All Science Journal Classification (ASJC) codes

  • Plant Science

Cite this

@article{333e62d41aa745f388127839b3dfe1df,
title = "A strategy for screening monoclonal antibodies for Arabidopsis flowers",
abstract = "The flower is one of the most complex structures of angiosperms and is essential for sexual reproduction. Current studies using molecular genetic tools have made great advances in understanding flower development. Due to the lack of available antibodies, studies investigating the localization of proteins required for flower development have been restricted to use commercial antibodies against known antigens such as GFP, YFP, and FLAG. Thus, knowledge about cellular structures in the floral organs is limited due to the scarcity of antibodies that can label cellular components. To generate monoclonal antibodies that can facilitate molecular studies of the flower, we constructed a library of monoclonal antibodies against antigenic proteins from Arabidopsis inflorescences and identified 61 monoclonal antibodies. Twenty-four of these monoclonal antibodies displayed a unique band in a western blot assay in at least one of the examined tissues. Distinct cellular distribution patterns of epitopes were detected by these 24 antibodies by immunofluorescence microscopy in a flower section. Subsequently, a combination of immunoprecipitation and mass spectrometry analysis identified potential targets for three of these antibodies. These results provide evidence for the generation of an antibody library using the total plant proteins as antigens. Using this method, the present study identified 61 monoclonal antibodies and 24 of them were efficiently detecting epitopes in both western blot experiments and immunofluorescence microscopy. These antibodies can be applied as informative cellular markers to study the biological mechanisms underlying floral development in plants.",
author = "Qian Shi and Lian Zhou and Yingxiang Wang and Hong Ma",
year = "2017",
month = "2",
day = "28",
doi = "10.3389/fpls.2017.00270",
language = "English (US)",
volume = "8",
journal = "Frontiers in Plant Science",
issn = "1664-462X",
publisher = "Frontiers Media S. A.",

}

A strategy for screening monoclonal antibodies for Arabidopsis flowers. / Shi, Qian; Zhou, Lian; Wang, Yingxiang; Ma, Hong.

In: Frontiers in Plant Science, Vol. 8, 270, 28.02.2017.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A strategy for screening monoclonal antibodies for Arabidopsis flowers

AU - Shi, Qian

AU - Zhou, Lian

AU - Wang, Yingxiang

AU - Ma, Hong

PY - 2017/2/28

Y1 - 2017/2/28

N2 - The flower is one of the most complex structures of angiosperms and is essential for sexual reproduction. Current studies using molecular genetic tools have made great advances in understanding flower development. Due to the lack of available antibodies, studies investigating the localization of proteins required for flower development have been restricted to use commercial antibodies against known antigens such as GFP, YFP, and FLAG. Thus, knowledge about cellular structures in the floral organs is limited due to the scarcity of antibodies that can label cellular components. To generate monoclonal antibodies that can facilitate molecular studies of the flower, we constructed a library of monoclonal antibodies against antigenic proteins from Arabidopsis inflorescences and identified 61 monoclonal antibodies. Twenty-four of these monoclonal antibodies displayed a unique band in a western blot assay in at least one of the examined tissues. Distinct cellular distribution patterns of epitopes were detected by these 24 antibodies by immunofluorescence microscopy in a flower section. Subsequently, a combination of immunoprecipitation and mass spectrometry analysis identified potential targets for three of these antibodies. These results provide evidence for the generation of an antibody library using the total plant proteins as antigens. Using this method, the present study identified 61 monoclonal antibodies and 24 of them were efficiently detecting epitopes in both western blot experiments and immunofluorescence microscopy. These antibodies can be applied as informative cellular markers to study the biological mechanisms underlying floral development in plants.

AB - The flower is one of the most complex structures of angiosperms and is essential for sexual reproduction. Current studies using molecular genetic tools have made great advances in understanding flower development. Due to the lack of available antibodies, studies investigating the localization of proteins required for flower development have been restricted to use commercial antibodies against known antigens such as GFP, YFP, and FLAG. Thus, knowledge about cellular structures in the floral organs is limited due to the scarcity of antibodies that can label cellular components. To generate monoclonal antibodies that can facilitate molecular studies of the flower, we constructed a library of monoclonal antibodies against antigenic proteins from Arabidopsis inflorescences and identified 61 monoclonal antibodies. Twenty-four of these monoclonal antibodies displayed a unique band in a western blot assay in at least one of the examined tissues. Distinct cellular distribution patterns of epitopes were detected by these 24 antibodies by immunofluorescence microscopy in a flower section. Subsequently, a combination of immunoprecipitation and mass spectrometry analysis identified potential targets for three of these antibodies. These results provide evidence for the generation of an antibody library using the total plant proteins as antigens. Using this method, the present study identified 61 monoclonal antibodies and 24 of them were efficiently detecting epitopes in both western blot experiments and immunofluorescence microscopy. These antibodies can be applied as informative cellular markers to study the biological mechanisms underlying floral development in plants.

UR - http://www.scopus.com/inward/record.url?scp=85014872462&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85014872462&partnerID=8YFLogxK

U2 - 10.3389/fpls.2017.00270

DO - 10.3389/fpls.2017.00270

M3 - Article

C2 - 28293248

AN - SCOPUS:85014872462

VL - 8

JO - Frontiers in Plant Science

JF - Frontiers in Plant Science

SN - 1664-462X

M1 - 270

ER -