Can SDS-PAGE Determine Hemoglobin's Molecular Weight?
Yes, SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) can be used to determine hemoglobin's molecular weight. This method separates proteins based on their molecular weight, allowing for accurate determination.
Procedure
1. Sample Preparation
Mix hemoglobin with SDS and a reducing agent (e.g., β-mercaptoethanol) to denature the protein and impart a uniform negative charge.
2. Gel Electrophoresis
Load the prepared sample onto a polyacrylamide gel.
Apply an electric current; proteins migrate inversely proportional to their molecular weights.
3. Staining and Visualization
After electrophoresis, stain the gel (commonly with Coomassie Brilliant Blue) to visualize protein bands.
4. Molecular Weight Determination
Compare the migration distance of hemoglobin to that of standard proteins with known molecular weights to estimate its molecular weight.
This technique is widely used due to its simplicity and reliability in determining protein molecular weights.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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