Overview
- Columns bind proteins of interest while endotoxins flow through
- Reduce endotoxin levels to less than 0.01 EU/µg of protein
- Proteins are desalted
- Greater than 95% protein recovery
- Concentrate protein samples and remove detergents at the same time
- Effectively remove a wide range of detergents including SDS, Triton® X-100, CHAPS, NP-40, and Tween 20
- Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Kits are designed for the rapid removal of endotoxins from previously purified proteins or peptides, with protein recoveries of > 95% being achieved. Endotoxins, also known as lipopolysaccharides, are cell-membrane components of Gram-negative bacteria such as E. coli. Endotoxins liberated by Gram-negative bacteria are frequent contaminations of protein solutions derived from bioprocesses. Due to the high toxicity of endotoxins in vivo and in vitro, their removal from protein preparations is often necessary prior to the use of the protein in downstream applications. These kits efficiently reduce endotoxin levels to ≤ 0.01 EU/mg of protein, using spin column chromatography based on Norgen’s proprietary resin as the separation matrix. These kits are highly efficient in removing many different salts commonly used in the laboratory including, but not limited to, MgCl2, NaCl, KCl, CaCl2, LiCl and CsCl. The purified protein samples can be used in a number of downstream applications including sequencing, cloning, and in vitro and in vivo introduction into cells and organisms for various purposes. The simultaneous removal of salts while concentrating a dilute protein solution makes the kit a convenient method for preparing proteins before running many downstream applications such as SDS-PAGE, isoelectric focusing, X-ray crystallography, NMR spectroscopy, mass spectroscopy and other applications.
Mini Kit
The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Mini Kit is designed for the rapid removal of endotoxins from up to 200 μg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.
Maxi Kit
The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Maxi Kit is designed for the rapid removal of endotoxins from up to 4 mg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.
Details
Supporting Data
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| Kit Specifications | |
| Maximum Protein Input | 200 μg |
| Maximum Column Volume Input | 600 μL |
| Molecular Weight of Recovered Proteins | No Molecular Weight cut-off |
| Final Endotoxin Levels | ≤ 0.01 EU/μg protein |
| Protein Recovery | 90-95% |
| % Detergent Removal | 90-95% |
| Detergents that can be Removed | Including Triton® X-100, CHAPS, NP-40 and Tween 20 |
| Minimum Elution Volume | 50 μL |
| Time to Complete 10 Purifications | 20 minutes |
Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years from the date of shipment.
| Component | Cat. 22800 (25 preps) | Cat. 22200 (4 preps) |
|---|---|---|
| Binding Buffer J | 8 mL | 8 mL |
| Binding Buffer N | 4 mL | 20 mL |
| Wash Solution M | 50 mL | 130 mL |
| Wash Solution CIP | 20 mL | 60 mL |
| Wash Solution N | 30 mL | 130 mL |
| Wash Solution NIP | 20 mL | 60 mL |
| Elution Buffer G | 6 mL | 20 mL |
| Endotoxin Removal Solution | 1.5 mL | 1.5 mL |
| Protein Neutralizer EF | 2 mL | 2 mL |
| Maxi Spin Columns (assembled with Collection Tubes) | – | 4 |
| Mini Spin Columns | 25 | – |
| Collection Tubes | 25 | – |
| Maxi Spin Columns (assembled with Collection Tubes) | 4 | |
| Elution Tubes (1.7 mL) | 25 | – |
| Elution Tubes (50 mL) | 4 | – |
| Product Insert | 1 | 1 |
