How Are Proteins Smaller than 10 kDa Typically Detected? Is Electrophoresis Applicable?
Detecting proteins with molecular weights below 10kDa requires specialized methodologies, among which electrophoresis is a notable technique.
1. Electrophoresis Techniques
While SDS-PAGE is widely used for protein detection, proteins under 10kDa may migrate rapidly through the gel, potentially exiting from the bottom. Tricine-SDS-PAGE is an adapted electrophoresis method employing Tricine as a buffer, offering improved resolution for proteins in this molecular weight range.
2. Mass Spectrometry
This method stands out for its sensitivity and accuracy in detecting small proteins. Techniques such as liquid chromatography-tandem mass spectrometry (LC-MSMS) allow for both qualitative and quantitative analyses of these proteins.
3. Immunodetection
For proteins with known characteristics, such as amino acid sequences or specific epitopes, immunodetection techniques including ELISA and Western blot are effective. Although these methods necessitate specific antibodies, they provide high sensitivity and specificity.
4. High-Performance Liquid Chromatography (HPLC)
HPLC is frequently employed to separate and detect small proteins. By selecting suitable chromatographic columns and mobile phases, proteins smaller than 10kDa can be efficiently separated and analyzed.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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