Biphasic 蛋白質體學層析管柱套件: SCX + C18



雙相毛細層析管柱的發明在蛋白質體學分析技術領域 上是一項重大的突破，雙相層析管柱的組成：前端是PepSil的強陽離子層析管柱、 後端是PepSil的C-18層析管柱；雙相的層析管柱再搭配獨特的pH緩衝溶液，可成功 解決鹽類干擾分析蛋白質與胜肽的問題，而且其操作方法容易。

Figure 2 is the chromatogram of a 2-D LC/MS/MS analysis of a tryptic digestion of a mouse liver lysate using CTI’s PepSil biphasic column with a Finnigan LTQTM ion trap. This two dimensional HPLC separation significantly increases the overall system peak capacity. Over 4000 proteins were identified in this single analysis.
Other advantages of using the pH elution include low MS background and the accurate prediction of peptide elution based on their pIs (see Fig 3).



PepSil Biphasic Columns – SCX + C18
Items
 Description Pore Size (A) Part Number
Column
 0.32x50 + 0.15x100 300 CTIBiphase150
Column
 0.15x50 + 0.075x100 300 CTIBiphase100
Buffer Kit pH buffer set   CTIBiphase Buffers
Biphase Kit Column + Buffers   CTIBiphase Kit

Application: New! Bi-phase

Caption
Fig.2 Protein Separation using Pepsil Biphase Columns
Column: Pepsil Biphase SCX + C18
P/N: CTIBiphase
Sample: Trypsin digested mouse liver proteome
Method: pH step elution using MS-compatible buffers ( CTI Buffer kit)



Comparison of the separation of liver lysate using pH elution and salt elution.
Total proteins identified at each pH gradient step.

  pH 3.0 pH 3.5 pH 4.0 pH 4.5 pH 5.0 pH 5.5 pH 6.0 pH 7.0 pH 8.0 Total
pH Elution 315 823 1452 1483 1337 1224 1216 1224 1229 4493
Salt Elution 27 123 151 1227 1148 764 668 332 227 2212

Figure 3. Comparison of pI profile of unique peptides eluted by IMDL and the database of peptides digested theoretically by trypsin.