Multiple ERβ antisera label in ERβ knockout and null mouse tissues

Melissa A. Snyder, Tereza Smejkalova, Paul M. Forlano, Catherine S. Woolley*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

69 Scopus citations

Abstract

In the process of characterizing a custom-made affinity-purified antiserum for estrogen receptor beta (ERβ), ck5912, we used a number of common tests for specificity of ck5912 along with that of 8 commercially available ERβ antisera: Affinity Bioreagents PA1-310B, Invitrogen D7N, Upstate 06-629, Santa Cruz H150, Y19, L20, 1531, and Abcam 9.88. We tested their recognition of recombinant ERβ (rERβ) versus rERα, ERβ versus ERα transfected into cell lines, as well as labeling in wildtype (WT) versus estrogen receptor beta knockout (βERKO) and null (ERβSTL-/L-) mouse ovary, hypothalamus, and hippocampus. To our surprise, we found that while most of these antisera passed some tests, giving the initial impression of specificity, western blot analysis showed that all of them recognized apparently identical protein bands in WT, βERKO and ERβSTL-/L- tissues. We share these results with the goal of helping other researchers avoid pitfalls in interpretation that could come from use of these ERβ antisera.

Original languageEnglish (US)
Pages (from-to)226-234
Number of pages9
JournalJournal of Neuroscience Methods
Volume188
Issue number2
DOIs
StatePublished - May 2010

Keywords

  • Antibody
  • ERβ
  • Estrogen receptor beta
  • Knockout mice
  • βERKO

ASJC Scopus subject areas

  • Neuroscience(all)

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