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Location: Electrophoresis > Applications > 2D Protein Analysis > Detection

Applications
1D Protein Analysis
SDS PAGE
IEF Separation
Detection: Western Blotting
Detection: Gel Staining
Imaging
2D Protein Analysis
Sample Preparation
1st Dimension IEF
2nd Dimension SDS PAGE
Detection
Silver staining
Staining gels with Coomassie™ Brilliant Blue
References
Related files
Imaging
Troubleshooting
Ettan DIGE system
Ettan DIGE system
Fragment Analysis
Principles
Applications
Separation
Detection and Imaging

Detection

Once electrophoresis is complete, the gel must be analyzed to obtain information on the position and quantity of each protein. Because most proteins are not directly visible, the gel must be processed to determine the location and amount of the separated proteins. The most common analytical procedure is staining. Proteins are usually stained with silver or Coomassie™ Blue. Once the gel is stained, it can be photographed or dried on a backing for a record of the position and intensity of each band and then analyzed. Most detection methods used for SDS gels can be applied to second-dimension gels.

Autoradiography and fluorography

Autoradiography and fluorography are the most sensitive detection methods. To employ these techniques, the sample must consist of protein radiolabeled in vivo using either 35S, 14C, 3H, or, in the case of phosphoproteins,32P. For autoradiographic detection, the gel is simply dried and exposed to X-ray film or a storage phosphor screen. Fluorography is a technique that provides extra sensitiv-ity by impregnating the gel in a scintillant such as PPO (2,4-diphenyloxazole) prior to drying.