Desalting is a critical step in processing samples that have been isolated in the presence of salts and other small molecules. Resin-based desalting is performed by allowing the small molecules to freely enter the resin pore and be retarded in their flow through the packed resin bed while the high molecular weight molecules like proteins in the sample are excluded from the resin and rapidly exit the column.
The spin column method eliminates cumbersome column preparation or equilibration, allowing multiple-sample processing in 7,000 Da.
Easy-to-use- no cumbersome column preparation or equilibration
Fast- no waiting for protein to emerge by gravity-flow
High protein recovery- low non-specific protein binding resin maximizes protein recovery
Protocol for Desalting or Removing Small Molecules
- Equilibrate Coral 96-well Desalt Spin Plates to room temperature
- Remove the sealing material from the bottom of the plate and place it on top of a wash plate.
- Remove the sealing material from the top of the desalt plate.
- Place the assembly into a centrifuge with 96-well plate-carrier rotor and centrifuged at 1,000×g for 2 minutes to remove the storage buffer. Discard the flow-through.
- Rinse the wash plate three times with deionized water, dry and save for future use.
- Stack the desalt plate on top of a sample collection plate (blue), aligning the alphanumeric indices on the plates.
- Apply sample (20-100µl) to the center of the resin bed. To expel the entire sample, carefully touch pipette tip to the resin. For 20 µl protein samples (>300 µg/ml), apply a 20 µl stacker of water or buffer on top of resin bed after the sample wash has fully absorbed to ensure maximal protein recovery.
- Centrifuge the plate assembly at 1,000×g for 2 minutes to collect the desalt sample. Discard the desalt plate or reserve it for further balancing purpose.
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|Overall Proteomics Flyer
|File size: 6.3MB
|Proteomics General Brochure
|File size: 4MB
|Coral Desalting Resin
|File size: 1.9MB
Automated tryptic digestion procedure for HPLC/MS/MS peptide mapping of immunoglobulin gamma antibodies in pharmaceutics.