Suitable for immunoblotting.
Many recombinant proteins have been engineered to express a short sequence derived from the V5 molecule known as V5-tag. This tag facilitates the detection, isolation and purification of the proteins. Monoclonal antibodies reacting specifically with V5 and conjugated to Cy3 may be useful in various immunotechniques, and also to identify the expression of a V5 tagged fusion protein in situ by immunofluorescence.
Monoclonal Anti-V5 (mouse IgG1 isotype) is derived from the V5 hybridoma, produced by fusion of mouse myeloma cells and splenocytes from BALB/c mice immunized with a synthetic peptide corresponding to amino acid residues GKPIPNPLLGLDST (95-108) of the P/V proteins of the Paramyxovirus SV5, conjugated to KLH. The conjugate is then extensively dialyzed to remove unbound Cy3 fluorophore. Monoclonal Anti-V5, Cy3 conjugate recognizes the V5 tag sequence on V5 tagged fusion proteins by immunocytochemistry.
Recombinant DNA technology enables the attachment of genes of interest to specific sequences or genes th at can provide μaffinity handlesμ designed to enable the selective identification and purification of the protein of interest. The addition of a V5 tag sequence (GKPIPNPLLGLDST) to a given gene creates a stable fusion product th at does not appear to interfere with the bioactivity of the protein, or with the biodistribution of the tagged product. Many recombinant proteins have been engineered to express a short sequence derived from the V5 from simian virus known as V5-tag. Anti-“tag” agarose conjugates may facilitate the detection, isolation and purification of these proteins.
