Thermo Scientific™ Pierce™ Peptide Desalting Spin Columns

Product Code.	Quantity	unitSize
15848521	50 Columns	Pack of 50
15808242	25 Columns	Pack of 25

Product Code. 15848521 Supplier Thermo Scientific™ Supplier No. 89851

Description

Pierce Peptide Desalting Spin Columns provide a convenient and reproducible way to desalt and remove contaminants from peptide samples prior to analysis by mass spectrometry.

Thermo Scientific Pierce Peptide Desalting Spin Columns are ready-to-use centrifuge spin columns that enable efficient desalting of peptide samples following enzymatic digestion. The polymer-based hydrophobic resin contained in each column provides excellent binding and recovery characteristics for peptide samples in preparation for mass spectrometry and other methods. Each spin column can bind from 5 μg to 5 mg of native and TMT-labeled peptides. The spin column format allows processing of multiple samples (10–300 μL each) in parallel in approximately 30 minutes.

Mass spectrometry is essential for studying biological compounds. However, many of the buffers and reagents used for sample preparation interfere with mass spectrometry analysis, leading to poor-quality spectra and reduced sensitivity. Pierce Peptide Desalting Spin Columns are designed to remove many of the interfering hydrophilic sample contaminants with minimal sample loss, thus enabling sensitive and comprehensive analysis of digested protein samples by mass spectrometry.

• Remove MS-interfering contaminants—significantly reduces signal suppression and improves sensitivity in mass spectrometry
• High binding capacity—hydrophobic polymer-based resin allows for recovery of peptide loads of native and TMT-labeled peptides from 5 μg to 5 mg
• Easy to use—resin is provided in single-use spin column format that fits common 2-mL tubes
• Compatible—resin is validated with a variety of complex samples, including TMT-labeled peptides, and is stable under extreme pH conditions

Specifications

• Product Type: Spin Desalting Column
• Product Line: Pierce
• Content And Storage: Store at 2–8°C.
• Shipping Condition: Room Temperature
• Final Product Type: Peptide
• Detection Method: Mass Spectrometry
• Format: Spin Column
• Quantity: 50 Columns

Frequently Asked Questions (FAQs)

How do I determine if my sample is compatible and ready for LC-MS?

Samples prepared using LC/MS grade reagents are suitable for LC-MS; however, particulates and other small molecules can all interfere with liquid chromatography separation and mass spectrometer source ionization. We recommend visual inspection of samples for particulate matter and using Pierce Peptide Desalting Spin Columns (Cat. Nos. 89851, 89852), Pierce C18 Spin Tips (Cat. No. 84850) or an in-line C18 trap column (https://www.thermofisher.com/order/catalog/product/164564-CMD) to remove non-volatile salts before MS analysis.

After TMT reagent labeling, I still see a lot of excess TMT tags in my sample. How should I remove it?

We offer EasyPep MS sample preparation kits (EasyPep Maxi Sample Prep Kit (Cat. No. A45734), EasyPep Mini MS Sample Prep Kit (Cat. No. A40006), EasyPep 96 MS Sample Prep Kit (Cat. No. A45733)) that contain wash buffers specifically formulated to clean up TMT-labeled protein digests. To remove excess TMT reagent from samples prepared using other sample preparation methods, we recommend using Pierce Peptide Desalting Spin Columns (Cat. Nos. 89851, 89852) with extra washes using 5% methanol. The Pierce High pH Reversed-Phase Peptide Fractionation Kit (Cat. No. 84868) can also be used to remove excess unreacted TMT tags before collecting fractions.

I'm seeing a peak in my mass spectrometry analysis results that seems to always be present in my system. What can I do?

The peak most likely represents PEG or a similar contaminant. Clean your MS system and/or perform sample clean-up using Pierce Peptide Desalting Spin Columns (Cat. Nos. 89851, 89852) or Pierce C18 Resin. In addition, ensure that LC-MS pre-blended solvents are being used (formic acid/water, formic acid/acetonitrile, etc.)

I cleaned up my peptide sample using C18 or desalting columns, but I lost most of my peptides. What went wrong?

Peptides do not bind well to reversed phase resins at neutral pH or in the presence of organic solvents (e.g., acetonitrile). Acidify protein digest samples using formic acid or trifluoroacetic acid (TFA) to pH <3 before desalting. Ensure that samples do not contain organic solvents before and after clean-up by drying them down using a SpeedVac concentrator or equivalent.

If I already removed small contaminants and detergents at the protein level, do I still need to desalt my peptides prior to mass spectrometry analysis?

Yes. We recommend performing additional cleanup after protein digestion to remove any residual salts or partially digested proteins using Pierce Peptide Desalting Spin Columns (Cat. Nos. 89851, 89852), Pierce C18 Spin Tips (Cat. No. 84850) or an in-line C18 trap column (https://www.thermofisher.com/order/catalog/product/164564-CMD).

Can I use protein desalting columns for cleanup of peptide samples?

No. Protein desalting columns (e.g., Zeba desalting columns) use size exclusion and have a molecular weight cut-off that is typically too high for peptide samples. Peptide desalting, on the other hand, uses reversed-phase chromatography resins (e.g., C18 resin) to bind peptides for salt removal during washing.

How effective is the salt removal using Pierce Peptide Desalting Spin Columns?

With Pierce Peptide Desalting Spin Columns, more than 90% of salt or labeling tags such as TMT tags can be removed.

What is the reproducibility of Pierce Peptide Desalting Spin Columns from sample to sample?

With Pierce Peptide Desalting Spin Columns, the reproducibility coefficient of variation (CV) is ± 20%.