Protein Purity Analysis (Gel Filtration/Reversed Phase Chromatography)
Molecular sieves are materials containing precise and uniform tiny pores, which can be used to adsorb gases or liquids. Molecular sieve HPLC, also known as size exclusion chromatography (SEC), allows small molecular weight compounds to enter the gel pores, resulting in longer retention times; large molecular weight compounds cannot enter the pores and are eluted earlier. The advantage of molecular sieve chromatography is that it can handle at least 1 nmol of target protein with good separation results, and the separation time is very short, generally within 3 hours, while the obtained chromatographic separation peaks are also smaller.
Protein molecular sieve SEC purity analysis
Advantages of protein purification by SEC:
1. Can perform buffer exchange and desalting.
2. Can separate similar species that are difficult to separate by other purification techniques (such as protein fragments and oligomers).
3. Compatible with various solvents.
4. Does not rely on any specific form of the protein for retention and elution.
Disadvantages of protein purification by SEC:
1. Separation effects heavily depend on the column packing quality.
2. Non-specific interactions between the protein and the medium can reduce resolution.
3. Low resolution for complex protein mixtures.
4. To ensure sufficient resolution, the sample load must be small. This is a problem for high concentration proteins obtained by precipitation.
Depending on the relative polarity of the mobile phase and the stationary phase, liquid chromatography is divided into normal phase chromatography and reverse phase chromatography. When the polarity of the mobile phase is greater than that of the stationary phase, it is called reverse phase chromatography. Reverse phase chromatography is characterized by high resolution and good reproducibility, making it a core technique in the field of protein and peptide analysis. For peptides and certain proteins with relatively simple molecular structures, RPLC is an efficient method for purification and preparation. Reverse phase chromatography HPLC is mainly used for the separation of small molecular weight proteins and protein fragments with a molecular weight of <20,000 Da.
Protein reverse phase liquid chromatography purity analysis
BaiTaiPike Bio is equipped with both molecular sieve and reverse phase liquid chromatography techniques for protein/peptide purification analysis, which can be used for the separation and determination of various protein/peptide samples, meeting the purity analysis needs of various protein/peptide samples.
Sample requirements:
The total concentration of protein or peptide should not be less than 0.5 μg/μl, and the volume should not be less than 50 μl. If the sample needs desalting or sample concentration, the composition of the buffer should be specified.
Chinese/English project report
In the technical report, BaiTaiPike will provide you with a detailed bilingual technical report in Chinese and English, which includes:
1. Experimental procedures (Chinese and English)
2. Relevant chromatographic parameters (Chinese and English)
3. Raw data
4. Protein purity analysis results
One-stop service for protein purity analysis based on molecular sieve and reverse phase chromatography
You only need to place an order and send the samples
BaiTaiPike's one-stop service completes: sample processing - on-machine analysis - data analysis - project report
