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Direct collection onto Zitex and PVDF for Edman sequencing
Elimination of polybrene
Burkhart, WA., Moyer, MB., Bodnar, WM., Everson, AM., Valladares, VG., Bailey, JM., & Crabb, JW. (Ed.) (1995). Direct collection onto Zitex and PVDF for Edman sequencing: Elimination of polybrene. In Techniques in Protein Chemistry (Vol. 6, pp. 169-176) https://doi.org/10.1016/S1080-8914(06)80023-2
Polybrene is used routinely as a carrier on many Edman sequencers to help with the sample washout problem; however, it requires preconditioning before application of sample and is a potential source of interference for mass spectrometric analysis. This chapter discusses the use of polybrene, evaluate the use of alternative membranes as sequencing supports, and improve the performance of PVDF with peptides. Zitex and PVDF perform equivalently with all the model peptides, except Tyr-bradykinin, and PVDF is the superior support for that particular peptide. The chapter also discusses several factors that contributed to improved performance, including (1) use of G104 Zitex, (2) use of the standard reaction cartridge, (3) gas-phase delivery of R3 (TFA) during cleavage, and (4) keeping ethyl acetate washes to a minimum. Teflon tape is found to give similar performance in electro blotting as PVDF, and proteins electro blotted to Teflon tape are amenable to N-terminal sequence analysis and in situ digestion. Because of its ease of handling, Zitex remains a choice for direct collection of peptides; however, Teflon tape promises to offer an alternative to PVDF for electro blotting with several advantages.