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查看更多产品信息 Pierce™ Polyacrylamide Spin Desalting Columns, 7K MWCO, 0.7 mL - FAQs (89849, 89862)
2 个常见问题解答
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.
Find additional tips, troubleshooting help, and resources within our Protein Dialysis, Desalting, and Concentration Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.