The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications

Abstract Specialized epitope tags are widely used for detecting, manipulating or purifying proteins, but often their versatility is limited. Here, we introduce the ALFA-tag, a rationally designed epitope tag that serves a remarkably broad spectrum of applications in life sciences while outperforming established tags like the HA-, FLAG®- or myc-tag. The ALFA-tag forms a small and stable α-helix that is functional irrespective of its position on the target protein in prokaryotic and eukaryotic hosts. We characterize a nanobody (NbALFA) binding ALFA-tagged proteins from native or fixed specimen with low picomolar affinity. It is ideally suited for super-resolution microscopy, immunoprecipitations and Western…

• DF
  Deutsche Forschungsgemeinschaft

  Awards: JU 2957/1-1, SFB 1032, DFG JU 2957/1-1
• C
  Cancerfonden
• M
  Max-Planck-Gesellschaft
• V
  Vetenskapsrådet

  Award: 2014-5667