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Thermo Scientific™ Pierce™ Polyacrylamide Spin Desalting Columns, 7K MWCO, 0.7 mL
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Description
Desalting and buffer exchange are two of the most widely used applications in resin filtration chromatography. Thermo Scientific Pierce Polyacrylamide Spin Desalting Columns, 7K MWCO (molecular-weight cutoff), are ready-to-use, disposable, gel-filtration columns for the separation of proteins and other macromolecules from low-molecular-weight buffer salts and reagents.
Pierce Polyacrylamide Spin Desalting Columns, 7K MWCO, are pre-packed with porous polyacrylamide beads and enable buffer-exchange and desalting of proteins and other macromolecular samples based on a molecular-weight cutoff (MWCO) of 7000 Da. Unlike other size-exclusion media, polyacrylamide resin is not subject to enzymatic degradation and will not serve as a nutrient for microbial growth. The resin is very hydrophilic, thereby minimizing undesirable binding interactions between the resin and sample molecules. Contamination with low molecular weight sugars (which may occur with crosslinked dextran resin) is not a concern when using polyacrylamide resin. Oxidizing agents can be removed without destroying the support. This resin is susceptible to hydrolysis of amide groups under extreme pH conditions, so an operating pH of 2–10 is recommended at room temperature. The polyacrylamide resin can also be autoclaved at pH 5.5–6.5 for 30 minutes at 120°C.
The columns use a resin with beads that have a wet diameter of 45 to 90 μM and are ideal for separating peptides and small macromolecules (>7 kDa) from buffer salts and other compounds (less than 1K MWCO).
Features of polyacrylamide resin include:
• Stable in water, salt solutions, organic solvents, and alkaline or acidic conditions
• Excellent flow properties
• Heat stable
Applications
• Removing salts from protein solutions
• Removing phenol from nucleic acid preparations
• Separating excess crosslinker from conjugate preparations
• Removing excess derivatizing agents from modified proteins
• Removing unreacted dye from fluorescent antibodies
• Removing free radiolabel from labeled proteins
• Buffer exchange
Specifications
Specifications
| Description | Pierce™ Polyacrylamide Spin Desalting Columns, 7K MWCO, 0.7 mL |
| Product Type | Spin Desalting Column |
| Content And Storage | Upon receipt store at 4°C. |
| Format | Spin Column |
| Purification Target | Buffer Exchange, Protein |
| Column Type | Size-exclusion, Polyacrylamide Resin |
| MWCO | 7.0 kDa |
| Product Line | Pierce |
| Quantity | 25 Columns |
| Sample Volume (Metric) | 30 to 120 μL |
Frequently Asked Questions (FAQs)
You may remove excess solvent and smaller moieties by centrifugation through a microconcentrator. This will concentrate your protein sample.
(1) Choose microconcentrator tube (available from a variety of commercial suppliers) with a protein cutoff smaller than the molecular weight of the protein in the sample. Check our Pierce Protein Concentrators PES.
(2) Add 1 µL of 20% w/v SDS to a dry microconcentrator tube (if sample does not already contain SDS).
(3) Slowly add sample (a few microliters at a time) to membrane until membrane is completely wet. Centrifuge to near (but not complete) dryness.
(4) If intention is to desalt sample or buffer exchange: Add ~50 µL water to microconcentrator and spin until nearly dry. Repeat buffer exchange. Sample will remain on membrane. Check our Zeba desalting proteins.
(5) Using a new collection tube, invert membrane and spin at low speed (1000 x g) to elute protein from membrane. Add 2X SDS-Sample Buffer containing 10 mM DTT to membrane: vortex, invert and spin. Final volume should be ~20 µL.
For protein samples, Pierce Polyacrylamide Spin Desalting Columns (Cat. No. 89849) can be used to remove salts and other small molecular weight contaminates. Acetone precipitation (https://tools.thermofisher.com/content/sfs/brochures/TR0049-Acetone-precipitation.pdf) is recommended for low protein amounts provided you have at least 10 µg of sample.
For peptide samples derived from protein digests, we recommend using Pierce Peptide Desalting Spin Columns (Cat. No. 89852), Pierce C18 Spin Tips (Cat. No. 84850) or an in-line C18 trap column (https://www.thermofisher.com/order/catalog/product/164564-CMD) for sample clean-up.
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