The Superdex 200 Increase 10/300 GL is a high-resolution gel filtration column designed for size exclusion chromatography. It offers rapid purification and analysis of proteins and biomolecules‚ combining high sample load capacity with excellent resolution‚ making it ideal for laboratory research and protein characterization.
Overview and Intended Use
The Superdex 200 Increase 10/300 GL is a high-performance gel filtration column designed for size exclusion chromatography (SEC). It is primarily intended for research use‚ focusing on protein purification‚ characterization‚ and analysis. The column is optimized for small-scale preparative purification and analytical applications‚ offering high resolution and efficiency. It is ideal for laboratories requiring reliable and quantifiable results in various chromatography workflows. The column is not intended for clinical or diagnostic purposes. Its design enables rapid separation and purification of biomolecules‚ making it a versatile tool for researchers in biochemistry‚ biotechnology‚ and related fields. It is part of the Superdex 200 Increase series‚ which includes columns with different bed dimensions to suit various experimental needs.
Key Features and Benefits
The Superdex 200 Increase 10/300 GL offers high-resolution separation and efficient processing for proteins and biomolecules. Its increased bed height improves resolution‚ while the high sample load capacity ensures excellent purification efficiency. Designed for small-scale applications‚ it is ideal for both preparative and analytical workflows. The column provides rapid separation with minimal buffer consumption‚ making it cost-effective and time-efficient. Its steep selectivity curve ensures precise fractionation of proteins within a wide molecular weight range. Additionally‚ the column is compatible with a wide range of buffers and pH conditions‚ enhancing its versatility for various chromatography needs. These features make it a reliable and efficient tool for protein purification and characterization in research settings.
Technical Specifications
The Superdex 200 Increase 10/300 GL has a bed height of 30 cm and diameter of 10 cm‚ with a bed volume of approximately 235 ml. It operates at pH 4–9‚ with a maximum pressure drop of 2.5 MPa and a sample load capacity of up to 10 mg.
Physical Dimensions and Bed Volume
The Superdex 200 Increase 10/300 GL column features a bed height of 30 cm and an inner diameter of 10 cm‚ providing an approximate bed volume of 235 ml. Its compact design ensures efficient separation and minimizes dead volume‚ making it ideal for small-scale protein purification and characterization. The column’s physical dimensions are optimized for high-resolution chromatography‚ allowing for precise separation of biomolecules with excellent recovery. The bed volume of 235 ml is sufficient for handling sample volumes up to 500 µl‚ ensuring optimal performance in analytical and preparative applications. Its glass construction ensures durability and chemical compatibility‚ making it a reliable choice for laboratory use.
Particle Size and pH Range
The Superdex 200 Increase 10/300 GL column features a median particle size of approximately 34 µm‚ ensuring high-resolution separation of biomolecules. The resin is designed to operate effectively within a pH range of 4.0 to 8.0 for standard use‚ allowing compatibility with a wide range of buffers commonly used in chromatography. For cleaning and sanitization‚ the pH range is slightly narrower‚ typically between 4.0 and 7;0‚ to maintain the integrity of the matrix. These specifications ensure optimal performance and longevity of the column‚ making it suitable for various protein purification and characterization applications. The particle size and pH tolerance are critical factors in achieving consistent and reliable chromatographic results.
Pressure Drop and Sample Load Capacity
The Superdex 200 Increase 10/300 GL column is designed to handle a maximum pressure drop of 2.5 MPa‚ ensuring compatibility with high-performance chromatography systems. It offers a high sample load capacity‚ accommodating up to 10 mg of protein in a 500 µl sample volume‚ making it ideal for both analytical and preparative applications. This capacity allows for efficient separation and purification of biomolecules without compromising resolution. The column’s design minimizes pressure-related issues while maintaining optimal flow rates‚ ensuring reliable performance across various experimental conditions. Its versatility in handling diverse sample loads makes it a valuable tool for laboratories requiring high-throughput protein purification and characterization.
Operating Instructions
- Unpack the column carefully‚ ensuring all components are intact.
- Prepare the sample by dissolving in eluent and filtering through a 0.22 µm filter.
- Connect the column to the chromatography system following the manual’s guidelines.
- Perform elution with the recommended flow rates to achieve optimal separation.
Unpacking and Installation
When unpacking the Superdex 200 Increase 10/300 GL column‚ ensure all components are undamaged and accounted for. Inspect the column and accessories for any visible defects. The column is typically supplied in a storage solution to maintain resin stability. Before installation‚ equilibrate the column by flushing it with the recommended buffer to remove any storage solution. Connect the column to your chromatography system according to the manufacturer’s instructions‚ ensuring compatibility with your equipment. For optimal performance‚ store the column at 4–30°C when not in use. Refer to the manual for detailed unpacking and installation procedures to ensure proper setup and functionality.
Sample Preparation and Column Connection
Before using the Superdex 200 Increase 10/300 GL column‚ prepare your sample by dissolving it in the appropriate eluent buffer. Filter the sample through a 0.22 µm filter or centrifuge at 10‚000 g for 10 minutes to remove particulates. Ensure the sample volume does not exceed the column’s recommended load capacity. Connect the column to your chromatography system‚ such as an FPLC‚ following the manufacturer’s guidelines. Use a flow restrictor (e.g.‚ FR-902) when disconnecting tubing at the injection valve to maintain system integrity. Properly equilibrate the column with the eluent before loading the sample to ensure optimal separation and resolution. Refer to the manual for specific instructions on connecting and preparing the column for operation.
Elution and Separation Principles
The Superdex 200 Increase 10/300 GL operates on size exclusion chromatography (SEC) principles‚ separating molecules based on their size. The column contains a bead-based matrix with defined pore sizes‚ allowing smaller molecules to penetrate the beads while larger molecules are excluded and elute earlier. Elution is typically performed with a continuous flow of buffer‚ ensuring consistent separation. The column’s design maximizes resolution and reduces cycle times‚ making it ideal for both research and preparative applications. The bead technology ensures high purity and recovery of target proteins‚ while maintaining their biological activity. This method is particularly effective for analyzing and purifying proteins‚ monoclonal antibodies‚ and other biomolecules‚ providing reliable and reproducible results.
Maintenance and Troubleshooting
Regular cleaning with 70% ethanol and storage at 4°C to 40°C are recommended. Avoid sodium azide for long-term storage if using with FPLC systems. Troubleshooting includes checking for column clogging or uneven flow‚ which can be resolved by washing with 1 M NaOH or replacing frits if necessary. Proper maintenance ensures optimal performance and longevity of the column.
Cleaning and Sanitization Procedures
For optimal performance‚ the Superdex 200 Increase 10/300 GL column should be cleaned and sanitized regularly. Start by flushing the column with 5 column volumes of 70% ethanol to remove impurities and sanitize. For more thorough cleaning‚ use 1 M NaOH solution‚ followed by extensive washing with distilled water to remove any residue. Avoid using sodium azide for long-term storage if the column is connected to an FPLC system. After cleaning‚ store the column in 20% ethanol at 4°C to 40°C to prevent microbial growth. Always handle the column gently to avoid damaging the packing. Proper cleaning ensures longevity and maintains chromatographic efficiency for consistent results in protein purification and analysis.
Storage Conditions and Shelf Life
The Superdex 200 Increase 10/300 GL column should be stored in a solution to maintain its performance and longevity. For short-term storage‚ keep the column in 20% ethanol at 4°C to 40°C to prevent microbial growth. Avoid using sodium azide as a preservative if the column is connected to an FPLC system. For long-term storage‚ ensure the column is tightly sealed and stored at the same temperature range. The shelf life is indefinite if stored properly under these conditions. Always handle the column with care to avoid damage to the bed or connectors. Proper storage ensures the column remains functional and ready for use in future chromatography applications. Follow the manufacturer’s guidelines for storage to maintain optimal performance.
Common Issues and Solutions
Common issues with the Superdex 200 Increase 10/300 GL include pressure drop fluctuations‚ inconsistent bed height‚ and sample precipitation. Pressure drop can be addressed by cleaning or replacing the column filter. Bed height inconsistencies may require recalibration or repacking the column. Sample precipitation can be mitigated by ensuring proper buffer compatibility and pH conditions. Regular cleaning and sanitization‚ as outlined in the manual‚ help prevent these issues. If problems persist‚ refer to the troubleshooting section of the official handbook or contact GE Healthcare support for assistance. Proper maintenance and adherence to operating guidelines ensure optimal column performance and longevity. Always follow the manufacturer’s recommendations to resolve issues effectively.
References and Further Reading
Refer to the GE Healthcare official manual for detailed instructions and troubleshooting. Additional resources are available on the GE Support website.
Official Manual and Handbook
The GE Healthcare official manual provides comprehensive guidance for the Superdex 200 Increase 10/300 GL‚ including detailed instructions for installation‚ operation‚ and maintenance. It covers critical aspects such as column unpacking‚ sample preparation‚ and elution principles. The manual also outlines safety precautions‚ storage conditions‚ and troubleshooting tips to ensure optimal performance. For advanced users‚ it includes sections on column efficiency‚ pressure drop calculations‚ and pH range considerations. Additionally‚ the handbook offers a deep dive into gel filtration principles‚ method development‚ and scalability for various applications. Refer to the official documentation for compliance with regulatory standards and to maximize the column’s lifespan and functionality.
Online Resources and Support
GE Healthcare provides extensive online resources for the Superdex 200 Increase 10/300 GL‚ including user manuals‚ troubleshooting guides‚ and technical specifications. The official GE Healthcare website offers a dedicated support portal with downloadable PDF manuals‚ FAQs‚ and knowledge base articles. Additionally‚ the GE Service & Support page provides access to customer service‚ repair requests‚ and training materials. Users can also find the Safety Data Sheets (SDS) and the GE Filtration Calibration Kit Manual for complementary tools. Online community forums and expert networks further assist with method optimization and troubleshooting. For comprehensive support‚ visit the official GE Healthcare resources and documentation library.