Bottom Up Proteomics - Peptide LCMS

 

High Resolution Peptide Columns

We recommend a 75u ID x 250 mm long column packed with 1.5u 100A Reprosil Saphir C18 particles for optimum resolution of complex peptide samples.  The LCMS base peak trace above shows the separation of 100 femtomoles of a BSA tryptic digest at a flow of 300 nl/min with a 50 minute gradient from 2-42% ACN in 0.1% formic acid.  For one dimensional separations of complex peptide mixtures, this column offers a good compromise between throughput and sensitivity to optimize resolution for maximum separation and identification of peptides (>95% coverage of BSA peptides in the example above).  Although one can use longer columns and/or longer run times to improve resolution for very complex samples (i.e. whole cell lysates), these samples are generally better handled using multidimensional separations (2DLC, Gel-LC, etc) due to LC pressure and MS utilization limits.  PremierLCMS also offers a wide range of high resolution peptide columns through our custom column packing services.

 

High Sensitivity Peptide Columns

 We recommend a 75u ID x 150 mm long column packed with 1.5u 100A Reprosil Saphir C18 particles for optimum sensitivity of low level peptide samples.  The LCMS traces above shows the separation and identification (LTQ-MS) of 50 attomoles of a BSA tryptic digest (>15% coverage of BSA peptides in the example above) at a flow of 300 nl/min with a 20 minute gradient from 2-42% ACN in 0.1% formic acid.  This column is well suited for many peptide applications including identification of 2D gel spot digests, trace protein digest peptide mapping, 2DLCMS of complex protein sample digests and peptidomics.  PremierLCMS also offers a wide range of high sensitivity peptide columns through our custom column packing services.

 

 Ultra High Throughput Peptide Columns

 We recommend a 150u ID by 50 mm long column packed with 1.5u 100A Reprosil Saphir C18 particles for optimum throughput of peptide samples.  The LCMS base peak trace above shows the separation of 100 femtomoles of a BSA tryptic digest at a flow of 2000 nl/min with a 2 minute gradient from 2-42% ACN in 0.1% formic acid. This column is well suited for many high throughput peptide applications including rapid quantitative proteomics, clinical proteomics and high throughput peptidomics.  PremierLCMS also offers a wide range of high throughput peptide columns through our custom column packing services.