Besides precipitation with ammonium sulfate, what are some ways I can concentrate my protein sample?
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.
What bucket adaptors are recommended for 20-100 mL capacity Pierce Protein Concentrators PES?
The dimensions of the 20-100 mL capacity concentrators (Pierce Protein Concentrators PES, 3K or 5K MWCO, 0.5–100 mL (Cat. No. 88534), Pierce Protein Concentrators PES, 10K MWCO, 0.5–100 mL (Cat. No. 88535), Pierce Protein Concentrators PES, 30K MWCO (Cat No. 88536), Pierce Protein Concentrators PES, 50K MWCO, 0.5–100 mL (Cat. No. 88542), and Pierce Protein Concentrators PES, 100K MWCO, 0.5–100 mL (Cat No. 88537)) are 61 mm outer diameter and 124 mm height. These are similar to Nalgene and Nunc 250 mL bottles. Adapters to the Nunc 250 mL bottles will fit to the 20-100 mL capacity concentrators.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
What are the dimensions of the Pierce Protein Concentrators PES, 3K or 5K MWCO, 0.5-100 mL (Cat. No. 88534)?
The dimensions of the Pierce Protein Concentrators PES, 3K or 5K MWCO, 0.5-100 mL (Cat. No. 88534) are as follows:
- Outer Diameter–61 mm
- Length–124 mm
Find additional tips, troubleshooting help, and resources within our Protein Dialysis, Desalting, and Concentration Support Center
What kind of rotor do you recommend using with a protein concentrator that can handle 20-100 mL sample volume?
We recommend using a rotor that can accommodate a 250 mL Nalgene bottle.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
Can I autoclave PES protein concentrators?
We do not recommend autoclaving the PES protein concentrators. High temperatures will significantly increase the membrane molecular weight cutoff (MWCO). To sterilize, we recommend using a 70% ethanol solution.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.