The Anti-Glucocorticoid Receptor Antibody Clone 5E4: Raising Awareness of Unspecific Antibody Binding

• Published in: International journal of molecular sciences Vol. 23; no. 9; p. 5049
• Main Authors: Ehlers, Lisa, Kirchner, Marieluise, Mertins, Philipp, Strehl, Cindy, Buttgereit, Frank, Gaber, Timo
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 02.05.2022; MDPI
• Subjects: AMP deaminase; Antibodies; antibody specificity; Binding; Binding sites; Cells, Cultured; Clone Cells - metabolism; Cloning; cross-reactivity; Experiments; Flow cytometry; glucocorticoid receptor; Glucocorticoids; Mass spectrometry; Medical research; Molecular weight; Peptides; Proteins; Reagents; Receptors, Glucocorticoid - metabolism; replication crisis; Scientific imaging; Therapeutic targets; Transcription Factors - metabolism; glucocorticoid receptor; cross-reactivity; antibody specificity; replication crisis
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23095049

Unspecific antibody binding takes a significant toll on researchers in the form of both the economic burden and the disappointed hopes of promising new therapeutic targets. Despite recent initiatives promoting antibody validation, a uniform approach addressing this issue has not yet been developed. Here, we demonstrate that the anti-glucocorticoid receptor (GR) antibody clone 5E4 predominantly targets two different proteins of approximately the same size, namely AMP deaminase 2 (AMPD2) and transcription intermediary factor 1-beta (TRIM28). This paper is intended to generate awareness of unspecific binding of well-established reagents and advocate the use of more rigorous verification methods to improve antibody quality in the future.