Abstract
The AI215 antibody against the EPEA tag recognizes an EPEA-tagged human TAC protein by immunofluorescence in paraformaldehyde-fixed HeLa cells.
Highlights
The EPEA tag is an extremely short epitope derived from the human α-Synuclein protein (Uniprot #P37840), used for detection and purification of tagged proteins with the NbSyn2 antibody (De Genst at al., 2010)
Materials & Methods Antibodies: The ABCD_AI215 antibody (ABCD nomenclature, web.expasy.org/abcd/; Lima et al, 2020) was produced by the Geneva Antibody Facility as a mini-antibody with the antigen-binding VHH fused to a mouse IgG2A Fc
The EPEA-tagged TAC protein is expected to be mostly present at the cell surface
Summary
The EPEA tag ( called C tag) is an extremely short epitope derived from the human α-Synuclein protein (Uniprot #P37840), used for detection and purification of tagged proteins with the NbSyn2 antibody (De Genst at al., 2010). HEK293 suspension cells (growing in FreeStyleTM 293 Expression Medium, Gibco #12338) were transiently transfected with the vector coding for the scFv-Fc. Supernatant (90 mg/L) was collected after 4 days. Antigen: HeLa cells (growing in DMEM GlutaMAXTM, Gibco #31966; supplemented with 8% Fetal Bovine Serum, Gibco #10270) cultured on glass coverslips (Menzel-Gläser, 22x22 mm) and transiently transfected 2 days before the experiment with an EPEA-tagged TAC protein (Uniprot #P01589), were used to detect the peptide tag.
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