Nanobodies against NanoTags now easier to access

Antibodies recognizing the VHH05-tag and 127D01-tag epitopes now commercially available

The nanobodies recognizing the VHH05-tag and 127D01-tag, two short epitope tags developed by our group for use in Drosophila, are now available in purified form from Cell Signaling Technology (CST).

These "NanoTags" and their corresponding high-affinity nanobodies (NbVHH05 and Nb127D01) were characterized for use in flies, including CRISPR-mediated knock-in tagging of endogenous genes, immunostaining, immunoblotting, and immunoprecipitation (Xu, Kim et al., 2022, eLife; Kim, Xu et al., 2022, Current Protocols; see also our earlier Light up your flies! post). 

Because the tags are short linear epitopes, they can be inserted at the N-terminus, C-terminus, or internally in a protein of interest with minimal disruption, and the resulting tagged protein can then be detected using the corresponding nanobody — as a conventional antibody for staining/blotting/IP, or expressed in vivo (e.g., as GFP-fusion "chromobodies") for live imaging and protein manipulation. Researchers interested in using this system can use CRISPR to knock-in either tag into an endogenous Drosophila gene, then use the matching nanobody to detect, localize, or manipulate the tagged protein. Tandem use of both NanoTags on a single secreted protein has also enabled sensitive, sandwich-format detection of circulating fly proteins (Han, Xia, Kim et al., 2026, PNAS).

Access to purified nanobodies was previously a potential barrier to use of this system by individual labs without specialized experience handling nanobodies. So, we wanted to make the community aware that purified nanobodies recognizing VHH05 and 127D01 are now commercially available.

  • VHH05-Tag (E5X7B) Alpaca VHH Monoclonal Antibody, #70643 — cellsignal.com
  • 127D01-Tag (E8J2W) Llama VHH Monoclonal Antibody, #44664 — cellsignal.com

Additional relevant resources are available at the Bloomington Drosophila Stock Center (BDSC), including these two fly stocks:

https://flybase.org/reports/FBst0094015

https://flybase.org/reports/FBst0094016

References
Xu, Kim et al. (2022) Protein visualization and manipulation in Drosophila through the use of epitope tags recognized by nanobodies. eLife. PubMed: 35076390
Kim, Xu et al. (2022) NanoTag Nanobody Tools for Drosophila In Vitro and In Vivo Studies. Current Protocols. PubMed: 36571722
Han, Xia, Kim et al. (2026) Phage display–mediated immuno-PCR to detect low-abundance secreted proteins in Drosophila. PNAS. PubMed: 41941621

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