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. However, 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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