Development and characterization of monoclonal antibodies to detect Klotho

Astha Maltare, Angela K. Nietz, Ann M. Laszczyk, Taylor S. Dunn, Mary E. Ballestas, Mary Ann Accavitti-Loper, Gwendalyn D. King

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Although antibodies are commercially available to allow investigation into the biology of the age-regulating protein Klotho, problems with antibody specificity and application functionality are significant barriers to progress. Chief among these limitations is the inability of current tools to allow in vivo validation of binding partners originally identified through transfection of tagged proteins. To overcome this barrier, we generated a series of hybridoma cell lines by immunizing rats with a GST-KL1 fusion protein. Purified antibodies generated from these cell lines differentially detect human or mouse Klotho protein via Western blot, immunocyto/histochemistry, and immunoprecipitation. Specificity of antibody binding to Klotho was confirmed by mass spectrometry following immunoprecipitation. With this confidence in antibody specificity, co-immunoprecipitation was utilized to validate the interaction of Klotho/FGFR and Klotho/wnt7a in mouse kidney lysates.

Original languageEnglish (US)
Pages (from-to)420-427
Number of pages8
JournalMonoclonal Antibodies in Immunodiagnosis and Immunotherapy
Volume33
Issue number6
DOIs
StatePublished - Dec 1 2014
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

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