Production and characterization of monoclonal antibodies to Xenopus proteins

Brett Horr; Ryan Kurtz; Ankit Pandey; Benjamin G. Hoffstrom; Elizabeth Schock; Carole LaBonne; Dominique Alfandari

doi:10.1242/dev.201309

Production and characterization of monoclonal antibodies to Xenopus proteins

Brett Horr
, Ryan Kurtz
, Ankit Pandey
, Benjamin G. Hoffstrom
, Elizabeth Schock
, Carole LaBonne
, Dominique Alfandari

Producción científica: Article › revisión exhaustiva

8 Citas (Scopus)

Resumen

Monoclonal antibodies are powerful and versatile tools that enable the study of proteins in diverse contexts. They are often utilized to assist with identification of subcellular localization and characterization of the function of target proteins of interest. However, because there can be considerable sequence diversity between orthologous proteins in Xenopus and mammals, antibodies produced against mouse or human proteins often do not recognize Xenopus counterparts. To address this issue, we refined existing mouse monoclonal antibody production protocols to generate antibodies against Xenopus proteins of interest. Here, we describe several approaches for the generation of useful mouse anti-Xenopus antibodies to multiple Xenopus proteins and their validation in various experimental approaches. These novel antibodies are now available to the research community through the Developmental Study Hybridoma Bank (DSHB).

Idioma original	English
Número de artículo	dev201309
Publicación	Development (Cambridge)
Volumen	150
N.º	4
DOI	https://doi.org/10.1242/dev.201309
Estado	Published - feb 2023

Nota bibliográfica

Publisher Copyright:
© 2023. Published by The Company of Biologists Ltd.

Financiación

This work was supported by a grant from the National Institutes of Health (R24OD021485 from the office of the directorate to D.A.). D.A. is also supported by grants from the National Institute of Dental and Craniofacial Research (R01DE016289 to D.A. and R01DE026434 to S. Moody and D.A.). Deposited in PMC for release after 12 months. We thank all Alfandari laboratory members for their discussion and suggestions; Dr Irini Topalidou and Dr Amy Burnside for proofreading the manuscript; Dr Inchul Yeo for his help in the characterization of the antibody to VegT; Dr Sergei Sokol, Takuya Nakayama and Helen Willsey for their help in the characterization of multiple monoclonal antibodies; all Xenopus contributors that provided plasmids to produce proteins; and the current and former staff at the Fred Hutch Antibody Technology Resource and the University of Massachusetts Amherst IALS microscopy and mass spectrometry facilities for their help. Mass spectral data were obtained at the University of Massachusetts Mass Spectrometry Core Facility (RRID: SCR_019063). Research reported in this publication was supported by the Office Of The Director, National Institutes Of Health of the National Institutes of Health under Award Number S10OD010645.

Financiadores	Número del financiador
National Institutes of Health (NIH)	S10OD010645, R24OD021485
National Institute of Dental and Craniofacial Research	R01DE016289, R01DE026434
Office of the Director
Pan-Massachusetts Challenge

ASJC Scopus subject areas

Molecular Biology
Developmental Biology

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abstract = "Monoclonal antibodies are powerful and versatile tools that enable the study of proteins in diverse contexts. They are often utilized to assist with identification of subcellular localization and characterization of the function of target proteins of interest. However, because there can be considerable sequence diversity between orthologous proteins in Xenopus and mammals, antibodies produced against mouse or human proteins often do not recognize Xenopus counterparts. To address this issue, we refined existing mouse monoclonal antibody production protocols to generate antibodies against Xenopus proteins of interest. Here, we describe several approaches for the generation of useful mouse anti-Xenopus antibodies to multiple Xenopus proteins and their validation in various experimental approaches. These novel antibodies are now available to the research community through the Developmental Study Hybridoma Bank (DSHB).",

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AU - Pandey, Ankit

AU - Hoffstrom, Benjamin G.

AU - Schock, Elizabeth

AU - LaBonne, Carole

AU - Alfandari, Dominique

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Production and characterization of monoclonal antibodies to Xenopus proteins

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Financiación

ASJC Scopus subject areas

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