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MtoZ Biolabs - Model pharm-purity-analysis-sds-page -SDS-PAGE Protein Purity Analysis Service
Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is a technology commonly used in protein purification and analysis. SDS-PAGE can separate proteins according to the differences in the charge and the different mobility due to different molecular sizes. If the protein sample has been highly purified and contains only one protein, the results would show a single protein band after SDS-PAGE separation. However, when there are multiple proteins in the protein samples, different proteins can be separated into multiple protein bands through SDS-PAGE. Therefore, SDS-PAGE technology provides a direct way to analyze the purity of protein samples. To meet the research needs of analyzing the purity of protein products, MtoZ Biolabs has developed and optimized a SDS-PAGE based workflow, and provides an accurate analytical service for protein/peptide purification study.
SDS is a type of anionic surfactant that can break the hydrogen and hydrophobic bonds in proteins. SDS can bind to proteins in a certain proportion to form SDS-protein complexes, covering the intrinsic charges of proteins. Therefore, the migration speed of all kinds of SDS-protein complexes is only determined by the molecular weight of proteins. Different proteins are separated by SDS-PAGE electrophoresis, followed by protein staining and analysis of protein bands.
Figure 1. SDS-PAGE Analytical Workflow
1. Determination of Protein Concentration
2. Sample Preparation
Add 2x loading buffer (5% β-ME) and boil sample for 10 minutes.
3. Electrophoretic Preparation
Gel production, installation of the electrophoretic tank, and preparation of electrophoretic buffer.
4. Protein Samples Loading
According to the protein concentration, take an appropriate amount of processed protein samples and add into the pocket of the gel. After loading all samples, add protein standard marker into the first or last pocket of the gel.
5. Electrophoresis
Turn on the power supply, adjust the voltage to 100V, and run the gel the constant voltage.
6. Gel Staining
At the end of the electrophoretic separation, staining with R250 stain for 1 hours, followed by destaining of protein gel until background become clean and clear.
7. Analysis of Purity and Molecular Weight of Protein Samples
1. Experiment Procedures
2. Parameters of SDS-PAGE
3. Protein Purity Results
4. Bioinformatics Analysis
MtoZ Biolabs is an integrate contract research organization (CRO) providing advanced proteomics, metabolomics, bioinformatics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. The name of MtoZ represents “mass to charge ratio” in mass spectrometry analysis, as most of our services are provided based on our well-established mass spectrometry platforms. Our services allow for the rapid and efficient development of research projects, including protein analysis, proteomics, and metabolomics programs.
MtoZ Biolabs is specialized in quantitative multiplexed proteomics and metabolomics applications through the establishment of state-of-the-art mass spectrometry platforms, coupled with high-performance liquid chromatography technology. We are committed to developing efficient, and effective tools for addressing core bioinformatics problems. With a continuing focus on quality, MtoZ Biolabs is well equipped to help you with your needs in proteomics, metabolomics, bioinformatics, and biopharmaceutical research. Our ultimate aim is to provide more rapid, high-throughput, and cost-effective analysis, with exceptional data quality and minimal sample.
Email: marketing@mtoz-biolabs.com