How to Interpret the Electrophoresis Results of SDS-PAGE for Target Protein Detection?
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is a widely used technique for protein separation. This method relies on sodium dodecyl sulfate (SDS) to denature proteins and confer a uniform negative charge, facilitating their separation in a polyacrylamide gel. Since SDS eliminates charge-based differences, protein migration is primarily dictated by molecular weight rather than the protein’s intrinsic charge or conformation.
Interpreting SDS-PAGE Electrophoresis Results
Following SDS-PAGE, the resulting gel displays multiple protein bands, each corresponding to a protein species. The position of a band (i.e., migration distance) provides an estimate of molecular weight, while its intensity and width reflect protein abundance.
Typically, higher molecular weight proteins migrate shorter distances, whereas lower molecular weight proteins travel further. By comparing band positions to molecular weight markers of known proteins, the approximate molecular weight of the target protein can be determined.
Identifying the Target Protein
If the expected molecular weight of the target protein is known, it can be identified by locating the corresponding band. However, proteins of similar molecular weights may co-migrate, making precise identification challenging.
To confirm the presence of the target protein, Western blotting can be employed using a specific antibody. This technique enables selective detection, thereby increasing confidence in protein identification.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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