2-d Gel Electrophoresis

Procedure

• First dimension: Isoelectric focusing (IEF) using immobilized pH gradients (separation of proteins according to difference in isoelectric points) 
• Second dimension: SDS-polyacrylamide gel electrophoresis (SDS-PAGE) (separation according to difference in molecular weight)
• Capacity: 20µg-1 mg of protein

IEF and SDS-PAGE are available as individual services as well as in combination.

Analysis Options

• pH ranges: 4-7 linear, 3-10 linear, 4-7 non-linear, 3-10 non-linear 
• SDS page gels: 10%, 12%, 4-12%gradient, 4-20% gradient, 10-20%gradient 
• Gel sizes: small: 7 cm IPG strip/ minigel Large: 13 cm IPG strip/
• Stains: Coomassie Blue compatible with in-gel digest, MS and sequencing. Silver Stain compatible with in-gel digest, MS and sequence analysis

We also transfer non-stained gels to pvdf for Edman sequencing.

For sample submission, please download the Protein Services Request Form. 

*FOR SERVICE FEES, please contact the facility at 217-333-4695 or email proteinsciences@uiuc.edu.

Sample Preparation

Careful sample preparation is the most critical parameter for a successful separation.

• Follow the facility's Guidelines for Sample Submission
• Remove contaminants that affect 2d results

2D gel of a bacterial protein extract

Peptide Synthesis and Purification | Protein Sequence Analysis | Mass Spectrometry