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PTMScan® Peptide Purification Kit
Proteomic Analysis Products

PTMScan® Peptide Purification Kit #35741

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To Purchase # 35741
Cat. # Size Qty. Price
1 Kit  (10 Purifications)
$ 115

Product Includes Volume (with Count)
PTMScan® C18 Reversed-Phase Columns 1 x 10 ea
PTMScan® Sample Reservoir Syringes 1 x 10 ea

Product Usage Information


• Approximately 20 mg of protease digested peptides can be purified on one C18 reversed-phase column.

• The steps described in the Directions for Use section use gravity feed. In cases where the flow rate has dramatically decreased, you may apply gentle pressure to the column using the syringe plunger.

• Each time a solution is applied to the column, an air bubble can form at the narrow junction between the syringe and the column inlet. This will decrease the flow rate considerably. The bubble should be removed by flushing with a gel loading tip fitted on a P200 pipette.

1. Acidify the peptides prior to loading by adding 1/20 volume of 20% TFA for a final concentration of 1% TFA.

2. Clarify the peptide sample by centrifuging at 1,780 x g at room temperature for 15 minutes.

3. Remove and save the plunger from a 10 ml syringe and connect to the short end of the C18 column.

4. Pre-wet the C18 reversed-phase column with 5 ml of 100% acetonitrile

5. Equilibrate the column sequentially with three washes of 0.1% TFA using 1, 5, and 6 ml.

6. Load the acidified peptide sample onto the column; the peptides will bind to the C18 material.

7. Wash the column sequentially with three washes of 0.1% TFA using 1, 5, and 6 ml.

8. Wash the column with 2 ml of 5% acetonitrile with 0.1% TFA.

9. Place each column above new, clean 50 mL collection tube.

10. Elute the peptides 3 times with 3 ml of 40% acetonitrile with 0.1% TFA.

11. Eluted peptides should then be frozen completely to proceed to the lyophilization step. Freeze for 4-16 hours in -80°C freezer or on a dry ice and ethanol bath.

See specific PTMScan protocol for further details.


Columns and syringes can be stored in the original sealed pouch for up to 24 months at room temperature. For best results, open pouches should be closed and sealed with tape, store resealed package in a desiccator for up to 6 months.

Product Description

C18 Reversed-phase solid phase extraction columns are useful tools for purifying and desalting peptides in preparation for PTMScan protocols and other mass spectrometry applications.


Reversed-phase chromatography is an important technique for analytical and preparative biochemical separation and purification. Hydrophobic proteins, peptides, and nucleic acids can be separated by reversed-phase chromatography with excellent resolution and recovery. Furthermore, the use of ion pairing modifiers in the mobile phase allows reversed-phase chromatography of charged solutes such as hydrophilic peptides and fully deprotected oligonucleotides. Preparative reversed-phase chromatography has been used in applications ranging from micro-purification of protein fragments for sequencing (1) to process scale purification of recombinant protein products (2) and mass spectrometry proteomic analysis (3).
  1. Renlund, S. et al. Peptides 14, 1125-32.
  2. Olson, C.V. et al. (1994) J Chromatogr A 675, 101-12.
  3. Rush, J. et al. (2005) Nat Biotechnol 23, 94-101.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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