superdex 200 increase 10/300 gl manual

Superdex 200 Increase 10/300 GL⁚ A Versatile Chromatography Column

The Superdex 200 Increase 10/300 GL is a prepacked column designed for high-resolution separation of biomolecules. It is ideal for separating proteins, peptides, and other biomolecules according to their size. This versatile column offers a wide separation range, making it suitable for various applications, including protein purification, analysis of aggregates, studying complex formation, and screening samples.

Introduction

The Superdex 200 Increase 10/300 GL is a versatile, prepacked chromatography column designed for high-resolution size exclusion chromatography (SEC) of proteins and other biomolecules. It is a valuable tool for researchers in various fields, including biochemistry, molecular biology, and drug discovery. This column offers a wide separation range, allowing for the separation of molecules with molecular weights from 10,000 to 600,000. Its high resolution and sample load capacity make it suitable for both analytical and preparative applications.

The Superdex 200 Increase 10/300 GL column utilizes a cross-linked dextran gel matrix with a controlled pore size distribution. This matrix provides excellent separation efficiency and high recovery rates for a wide range of biomolecules. The column is designed for use with a variety of chromatography systems, including high-performance liquid chromatography (HPLC) and fast protein liquid chromatography (FPLC).

This manual provides comprehensive information on the Superdex 200 Increase 10/300 GL column, including its applications, key features and benefits, specifications, installation, operation, maintenance, troubleshooting, and comparison with other Superdex columns. By understanding the principles and procedures outlined in this manual, researchers can effectively utilize this column for their protein purification and analysis needs.

Applications of Superdex 200 Increase 10/300 GL

The Superdex 200 Increase 10/300 GL column finds broad application in various research and industrial settings due to its versatility and high performance in size exclusion chromatography. Its key applications include⁚

• Protein Purification⁚ The column effectively separates proteins based on their size, enabling the purification of target proteins from complex mixtures. This is particularly useful for isolating proteins for further analysis or functional studies.
• Analysis of Aggregates⁚ The Superdex 200 Increase 10/300 GL column can be used to analyze the presence and size distribution of protein aggregates. This is crucial for ensuring the quality and stability of therapeutic proteins, particularly monoclonal antibodies.
• Studying Protein-Protein Interactions⁚ By separating protein complexes based on their size, the column helps in studying the formation and stability of protein interactions. This information is valuable for understanding biological processes and developing new therapeutic targets.
• Screening Samples⁚ The column can be used to quickly screen samples for the presence of specific proteins or biomolecules. This is useful in drug discovery, where it can help identify lead compounds that interact with target proteins.
• Polishing Step in Purification Procedures⁚ The Superdex 200 Increase 10/300 GL column can be used as a final polishing step in purification procedures to remove any remaining contaminants or aggregates. This ensures the purity and integrity of the final product.

The versatility of the Superdex 200 Increase 10/300 GL column makes it an indispensable tool for researchers working with proteins and other biomolecules.

Key Features and Benefits

The Superdex 200 Increase 10/300 GL column boasts several key features and benefits that make it a valuable tool for protein purification and analysis⁚

• High Resolution⁚ The column provides high-resolution separations, allowing for the precise separation of proteins and other biomolecules based on their size. This ensures accurate identification and analysis of different components in a mixture.
• High Sample Load Capacity⁚ The Superdex 200 Increase 10/300 GL column can handle a significant amount of sample, making it suitable for both small-scale and preparative purifications. This translates to increased efficiency and productivity in your research or production workflow.
• Wide Separation Range⁚ The column offers a broad separation range for molecules with molecular weights between 10,000 and 600,000. This versatility allows for the analysis and purification of a wide variety of proteins and other biomolecules, making it adaptable to diverse research applications.
• Ease of Use⁚ The Superdex 200 Increase 10/300 GL column is pre-packed, simplifying its setup and operation. The column comes with a storage/transport device to prevent drying out and is pre-equilibrated with degassed 20% ethanol, further simplifying the process. This makes it user-friendly for researchers of all experience levels.
• Durability⁚ The column is designed for long-term use, ensuring its reliability and consistent performance over time. This translates to a significant return on investment, as the column can be used for numerous experiments and purifications.

These features and benefits make the Superdex 200 Increase 10/300 GL column a powerful tool for researchers, biochemists, and biopharmaceutical professionals who require high-quality protein purification and analysis.

Specifications and Ordering Information

The Superdex 200 Increase 10/300 GL column is a high-performance chromatography column designed for size exclusion chromatography of proteins and other biomolecules. Here are the key specifications and ordering information for this product⁚

• Column Dimensions⁚ 10 mm internal diameter x 300 mm length
• Separation Range⁚ 10,000 to 600,000 Da (globular proteins)
• Packing Material⁚ Superdex 200 Increase resin
• Flow Rate⁚ 0.5 to 1.0 mL/min
• Sample Volume⁚ Up to 500 μL
• Pressure Limit⁚ 1.5 MPa (220 psi)
• Temperature Range⁚ 4°C to 40°C
• Storage⁚ Store in 20% ethanol at 4°C

To order the Superdex 200 Increase 10/300 GL column, you can contact Cytiva directly or through authorized distributors. Be sure to provide the following information when placing your order⁚

• Product Code⁚ 17-5175-01
• Quantity⁚ The number of columns you require
• Shipping Address⁚ Ensure accurate and complete shipping details for timely delivery.

For more detailed information on specifications and ordering, please refer to the Cytiva website or contact their customer service team.

Installation and Operation

Installing and operating the Superdex 200 Increase 10/300 GL column is straightforward, but it’s essential to follow the recommended procedures to ensure optimal performance and longevity of the column. Here’s a step-by-step guide⁚

1. Unpacking and Inspection⁚ Carefully unpack the column and inspect for any damage or defects. Ensure the column is properly sealed and the packing material is intact.
2. Equilibration⁚ Before use, equilibrate the column with the desired running buffer. This ensures that the column is at the appropriate pH and ionic strength for your experiment;
3. Connecting the Column⁚ Connect the column to your chromatography system using appropriate tubing and fittings. Ensure tight connections to prevent leaks.
4. Loading the Sample⁚ Apply your sample to the column using a syringe or pump. The sample volume should be within the recommended range for the column.
5. Elution⁚ Elute the sample with the running buffer at the desired flow rate. The elution profile will depend on the size of the molecules in your sample.
6. Collecting Fractions⁚ Collect fractions of the eluent using a fraction collector or manually. The number and size of fractions will depend on your experimental design.
7. Cleaning and Storage⁚ After use, clean the column with a suitable cleaning solution according to the manufacturer’s instructions. Store the column in 20% ethanol at 4°C when not in use.

For more detailed instructions on installation and operation, refer to the Cytiva manual for the Superdex 200 Increase 10/300 GL column.

Maintenance and Troubleshooting

Maintaining the Superdex 200 Increase 10/300 GL column is crucial for ensuring its optimal performance and extending its lifespan. Regular cleaning and proper handling are essential to prevent clogging and degradation of the gel matrix. Here’s a guide to maintenance and troubleshooting⁚

1. Regular Cleaning⁚ After each use, flush the column with a suitable cleaning solution. This helps remove any residual sample components and contaminants. The recommended cleaning solution will depend on the specific application and sample type.
2. Filter Replacement⁚ The column has a filter at the top to prevent particulate matter from entering the gel bed. Regularly inspect the filter and replace it when necessary. A filter replacement kit is available from Cytiva.
3. High Back Pressure⁚ If the column exhibits high back pressure, it could be due to clogging, particle accumulation, or a damaged filter. Examine the column for any visible blockage and consider replacing the filter.
4. Reduced Resolution⁚ Decreased resolution could indicate degradation of the gel matrix or a faulty column packing. If the column is properly cleaned and the filter is replaced, contact Cytiva for further troubleshooting.
5. Abnormal Elution Profile⁚ If the elution profile deviates significantly from the expected pattern, it may suggest problems with the column or the chromatography system. Inspect the system for leaks or malfunctions.

For comprehensive troubleshooting guidance and detailed cleaning procedures, refer to the Cytiva manual for the Superdex 200 Increase 10/300 GL column.

Comparison with Other Superdex Columns

The Superdex 200 Increase 10/300 GL is part of a family of Superdex columns designed for size exclusion chromatography. Choosing the right Superdex column depends on the specific application and the size range of the biomolecules you are separating. Here’s a comparison with other Superdex columns⁚

• Superdex 200 Increase 5/150 GL⁚ This column is smaller than the 10/300 GL and is ideal for rapid purity checks and screening of proteins by size exclusion chromatography. It offers a shorter run time and requires a smaller sample volume, making it suitable for quick analysis.
• Superdex 75 Increase⁚ This column is designed for rapid purity checks and screening of proteins with molecular weights less than or equal to 44,000. It is a smaller column with a shorter run time, making it suitable for high-throughput applications.
• Superdex 30 Increase 32300⁚ This column is designed for high-resolution separation of biomolecules in the size range of 1,000 to 30,000 Da. It is suitable for applications like peptide analysis and small protein separations.

The selection of the appropriate Superdex column depends on the specific application, the size range of the target molecules, and the desired resolution. The Superdex 200 Increase 10/300 GL offers a balance of resolution, sample capacity, and run time, making it a versatile choice for many applications.

Superdex 200 Increase 10/300 GL⁚ A Valuable Tool for Protein Purification and Analysis

The Superdex 200 Increase 10/300 GL is a powerful tool for protein purification and analysis. Its high resolution and versatility make it suitable for a wide range of applications, including⁚

• Protein Purification⁚ The Superdex 200 Increase 10/300 GL can be used as a polishing step in a purification procedure, removing contaminants and obtaining highly purified protein samples.
• Analysis of Protein Aggregates⁚ The column can separate different protein aggregates based on their size, providing insights into the aggregation state of protein samples.
• Studying Protein-Protein Interactions⁚ The column can be used to study the formation of protein complexes by separating different protein species based on their size and interactions.
• Screening Samples⁚ The column can be used to screen samples for the presence of specific proteins or to analyze the size distribution of proteins in complex mixtures.

The Superdex 200 Increase 10/300 GL provides a convenient and reliable method for purifying and analyzing proteins, contributing to research in various fields, including biotechnology, pharmaceuticals, and diagnostics.

The Superdex 200 Increase 10/300 GL is a highly versatile and valuable tool for protein purification and analysis. Its high resolution, wide separation range, and compatibility with various buffer systems make it an indispensable tool in various research settings. This column provides a reliable and efficient method for separating proteins, peptides, and other biomolecules based on size, allowing researchers to purify proteins, analyze aggregates, study protein-protein interactions, and screen samples for specific proteins. The Superdex 200 Increase 10/300 GL serves as a powerful tool for advancing research in various fields, including biotechnology, pharmaceuticals, and diagnostics, contributing to the development of new therapies and treatments.

Whether you are working on a small-scale preparative purification or high-resolution analysis, the Superdex 200 Increase 10/300 GL provides the performance and reliability you need. Its user-friendly design and comprehensive documentation make it a valuable asset for any laboratory involved in protein research.

References

Cytiva Life Sciences. (2024). Superdex 200 Increase 10/300 GL⁚ Column for high-resolution purification and analysis of mAbs and proteins. [Product information sheet]. Retrieved from https://cdn.cytivalifesciences.com/dmm3bwsv3/AssetStream.aspx?mediaformatid=10061&destinationid=10016&assetid=17491

GE Healthcare. (2024). Superdex 200 Increase 10/300 GL. [Product information sheet]. Retrieved from https://www.gehealthcare.com/products/lifesciences/chromatography/size-exclusion-chromatography/superdex-200-increase-10-300-gl

NCBI. (2011). Structure of the C-terminal domain of Saccharomyces cerevisiae Nup133, a component of the Nuclear Pore Complex. [Research article]. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3350809/

Cytiva Life Sciences. (2024). Gel filtration, Principles & Methods. [Handbook]. Retrieved from https://www.cytivalifesciences.com/en/us/shop/chromatography/gel-filtration/gel-filtration-principles-and-methods