CTI's biphasic capillary column, which was introduced in 2005, is a revolutionary approach to proteomics analysis. This column combines PepSil SCX and C18 materials in a single column, making it ideal for high performance multidimensional LC/MS separations on single dimensional LC/MS systems. The first 5cm of the column contains PepSil SCX material and the remaining column length is filled with PepSil C18 material. Common problems with salt-elution of peptides from such a system are avoided with the use of MS-compatible pH steps available in a convenient, easy to use kit from CTI. 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.
| PepSil Biphasic Columns – SCX + C18 | ||||
| Items | Description | Pore Size (A) | Part Number | Price |
|---|---|---|---|---|
| Column | 0.32x50 + 0.15x100 | 300 | CTIBiphase320 | $1600.00 |
| Column | 0.15x50 + 0.075x100 | 300 | CTIBiphase150 | $1600.00 |
| Buffer Kit | pH buffer set | CTIBiphase Buffers | $500.00 | |
| Biphase Kit 1 | CTIBiphase320 + buffer set | CTIBiphase kit 1 | $2000.00 | |
| Biphase Kit 2 | CTIBiphase150 + buffer set | CTIBiphase kit 2 | $2000.00 | |
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)
Mouse liver protein found using biphasic column and 2D LC/MS/MS method.