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. Author manuscript; available in PMC: 2020 Oct 1.
Published in final edited form as: Mol Immunol. 2019 Sep 10;114:513–523. doi: 10.1016/j.molimm.2019.08.008

Figure 5:

Figure 5:

Comparison of immunoprecipitation properties of VHHs that recognize linear epitope tags. All VHHs besides VHH05 were previously reported25–27. The identity of the GFP tags are: 1- GG (no tag), 2-QADQEAKELARQIS (UBC6e 14-mer), 3- PDRKAAVSHWQQ (BC2 tag), 4-EPEA (EPEA tag). GFP tags were appended to the C-terminus of eGFP using sortagging with identity verified using LC/MS (data not shown). VHHs biotinylated using sortagging were used to immunoprecipitate tagged GFPs. The same preparation of tagged GFP was used for all of the immunoprecipitations with each of the different VHHs. VHHs and GFPs were incubated at equivalent concentrations (5 μM) in TBS + 10% glycerol followed by addition of streptavidin sepharose beads. Beads were washed with TBS + tween 20 and the immunoprecipitate was analyzed using SDS-PAGE on 15% polyacrylamide gels visualized with Coomassie staining.