TAILIEUCHUNG - Proteomic Applications in Biology Part 4

Tham khảo tài liệu 'proteomic applications in biology part 4', khoa học tự nhiên, công nghệ sinh học phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | 40 Proteomic Applications in Biology phosphines have gained widespread use in 2-DE being dithiothreitol DTT the most often used. Since DTT is charged especially at alkaline pH during IEF it will migrate out of the gel with a concomitant loss of solubility for some proteins and 2-DE horizontal streaking Herbert 1999 Molloy 2000 . Therefore its use in combination with other reducers or its substitution by compounds such as tributyl phosphine TBP Tris 2-carboxyethyl phosphine hydrochloride TCEP-HCP and hydroxyethyl disulfide HED is advisable Méchin et al. 2003 Sarma et al. 2008 Acín et al. 2009 Zhang et al. 2011 . Another advantage of using phosphines is the possibility of shortening the length of the equilibration step therefore diminishing the loss of proteins at this point Zuo Speicher 2000 . This can be accomplished by performing the reducing and alkylating procedures at the same time since the phosphines such as TBP do not react with alkylating agents such as acrylamide and 2-vinylpyridine Molloy 2000 . Salt ions help stabilize proteins therefore their absence may lead to protein precipitation. One way to overcome this situation is to add ampholytes to the IEF solution. These molecules enhance solubility of individual proteins as they approach their pI. They also buffer changes in conductivity scavenge cyanate derived from urea prevent interactions between hydrophobic proteins and IEF matrix and assist nucleic acids precipitation during centrifugation Shaw and Riederer 2003 Khoudoli et al. 2004 Gorg et al. 2009 Rabilloud Lelong 2011 . As expected improvements in the composition of the IEF solubilisation buffer should help overcome some of the problems mentioned earlier in this chapter. For instance the use of a reducing and an alkylating agent TBP and 2-vinylpyridine dissolved in a strong chaotrope such as guanidine hydrochloride to resuspend a dry fruit . peanut pegs protein pellet obtained after TCA acetone washes and phenol-based precipitation have improved

TÀI LIỆU LIÊN QUAN
TỪ KHÓA LIÊN QUAN
Đã phát hiện trình chặn quảng cáo AdBlock
Trang web này phụ thuộc vào doanh thu từ số lần hiển thị quảng cáo để tồn tại. Vui lòng tắt trình chặn quảng cáo của bạn hoặc tạm dừng tính năng chặn quảng cáo cho trang web này.