2A peptides and 2A-like peptide sequences (known as cis-acting hydrolase elements or CHYSEL) are a superior alternative to internal ribosomal entry sites (IRES) for coordinating the expression of multiple gene products from a single recombinant construct. The 2A sequences are relatively short peptides of about 20 amino acids (depending on the virus of origin) containing the consensus motif Asp-Val/Ile-Glu-X-Asn-Pro-Gly-Pro. 2A peptides allow multiple proteins to be encoded as polyproteins, which then dissociate into component proteins upon translation. The 2A sequence impairs normal peptide bond formation via a mechanism called ribosomal skipping, which results in effective, non-enzymatic generation of distinct peptide products from a single multicistronic construct. 2A peptides are used by several families of viruses, the best known foot-and-mouth disease virus of the Picornaviridae family, for producing multiple polypeptides. Anti-2A peptide, clone 3H4 recognizes 2A sequence derived from foot and mouth piconavirus (VKQTLNFDLLKLAGDVESNPG*P) with cleavage site between G and P. (Ref.: Trichas, G, et al. (2008). BMC Biol. 6: 40).
Application: Western Blotting Analysis: A representative lot detected 2A peptide in HEK293 SpCas9-P2A-Puro and NIH3T3 SpCas9-T2A-Puro cell lysates (Courtesy of Stefan Schuchner, Ph.D. and Egon Ogris, M.D., Medical University of Vienna, Austria).Immunoprecipitation Analysis: A representative lot detected 2A peptide in HeLa cells expressing SpCas9-P2A or plain HeLa cells, HEK293 cells expressing SpCas9-T2A or plain HEK293 cells (Courtesy of Stefan Schuchner, Ph.D. and Egon Ogris, M.D., Medical University of Vienna, Austria).Immunofluorescence Analysis: A representative lot detected 2A peptide in HeLa cells transiently transfected with SaCas9-T2A-GFP (Courtesy of Stefan Schuchner, Ph.D. and Egon Ogris, M.D., Medical University of Vienna, Austria).
Other Notes: Concentration: Please refer to lot specific datasheet.