Epitope mapping of commercial antibodies that detect myocilin

Athéna C. Patterson-Orazem, Shannon E. Hill, Michael P Fautsch, Raquel L. Lieberman

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The presence of myocilin is often used in the process of validating trabecular meshwork (TM) cells and eye tissues, but the antibody reagents used for detection are poorly characterized. Indeed, for over a century, researchers have been using antibodies to track proteins of interest in a variety of biological contexts, but many antibodies remain ill-defined at the molecular level and in their target epitope. Such issues have prompted efforts from major funding agencies to validate reagents and combat reproducibility issues across biomedical sciences. Here we characterize the epitopes recognized by four commercial myocilin antibodies, aided by structurally and biochemically characterized myocilin fragments. All four antibodies recognize enriched myocilin secreted from human TM cell media. The detection of myocilin fragments by ELISA and Western blot reveal a variety of epitopes across the myocilin polypeptide chain. A more precise understanding of myocilin antibody targets, including conformational specificity, should aid the community in standardizing protocols across laboratories and in turn, lead to a better understanding of eye physiology and disease.

Original languageEnglish (US)
Pages (from-to)109-112
Number of pages4
JournalExperimental Eye Research
Volume173
DOIs
StatePublished - Aug 1 2018

Fingerprint

Epitope Mapping
Antibodies
Trabecular Meshwork
Epitopes
Ocular Physiological Phenomena
Eye Diseases
trabecular meshwork-induced glucocorticoid response protein
Western Blotting
Enzyme-Linked Immunosorbent Assay
Research Personnel
Peptides

Keywords

  • Antibodies
  • Antigen
  • ELISA
  • Epitopes
  • Glaucoma
  • Myocilin
  • Trabecular meshwork
  • Western blot

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Epitope mapping of commercial antibodies that detect myocilin. / Patterson-Orazem, Athéna C.; Hill, Shannon E.; Fautsch, Michael P; Lieberman, Raquel L.

In: Experimental Eye Research, Vol. 173, 01.08.2018, p. 109-112.

Research output: Contribution to journalArticle

Patterson-Orazem, Athéna C. ; Hill, Shannon E. ; Fautsch, Michael P ; Lieberman, Raquel L. / Epitope mapping of commercial antibodies that detect myocilin. In: Experimental Eye Research. 2018 ; Vol. 173. pp. 109-112.
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