Sample Preparation Products

For rapid and efficient endotoxin removal from proteins and peptides Rapid and convenient spin column format Columns bind proteins of interest while endotoxins flow-through Reduce endotoxin levels to less than 0.01 EU/µg of protein Greater than 95% protein recovery

The ProteoSpin™ Endotoxin Removal Kit efficiently reduces endotoxin levels to ≤ 0.01 EU/µg of protein using spin column chromatography.   This kit works by binding proteins to the spin column matrix while endotoxins and other contaminants are washed away.  The purified proteins and peptides are then eluted and ready for downstream applications.  A unique feature of the kit is that the endotoxins can be washed away repeatedly until desirable levels of endotoxins are obtained.  Other commercially available kits employ membranes to bind the endotoxins and therefore can quickly become saturated with endotoxin, however Bio-Synthesis's columns do not become saturated as the endotoxins are not bound on the column.  In addition, no further purification steps are required with Bio-Synthesis's kit  to clean-up the sample prior to use in downstream applications. Bio-Synthesis's kit will desalt, remove detergents and allow for buffer exhange while simultaneously removing endotoxins, therefore saving considerable time and expense. The purified protein is suitable for in vivo and in vitro introduction into cells and organisms, monoclonal and polyclonal antibody generation, and other applications such as mass spectrometry, gel electrophoresis and immunoassays and Western blots. Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, Binding Buffer is added to the protein sample and the sample is placed onto the top reservoir of a column. Endotoxin Removal Solution is added, followed by the addition of isopropanol, and the sample is then spun through the column (BIND). Under these conditions only the proteins will bind to the column while endotoxins and other contaminants will be removed in the flowthrough. The bound proteins are then washed to further remove the endotoxins (WASH). Lastly, the endotoxin-free proteins are eluted into a small volume of the provided Elution Buffer (ELUTE). Please see the procedure flowchart to the right.

FEATURES AND BENEFITS Endotoxin-free proteins and peptides - The ProteoSpin™ Endotoxin Removal Micro Kit reduces endotoxin levels 0.01 EU/µg of protein or less. Fast and easy processing - Efficient processing of multiple samples in 20 to 30 minutes using an easy-to-use protocol. High recovery - Protein recovery is over 95% of input. No further clean-up of protein samples required - Bio-Synthesis's kit provides desalting, buffer exchange and detergent removal while removing endotoxins Recovered protein is suitable for downstream applications - Purified protein is suitable for in vivo and in vitro introduction into cells and organisms, monoclonal and polyclonal antibody generation, mass spectrometry, gel electrophoresis, immunoassays, and Western blots Columns are provided ready-to-use - Unlike other endotoxin removal methods, there is no need for lengthy column or resin preparation. No endotoxin carry-over - No column regeneration, unlike other gel-based endotoxin removal procedures, which can lead to endotoxin carry-over. Columns are single use only.

APPLICATIONS Proteins and peptides purified using Bio-Synthesis's ProteoSpinTM Endotoxin Removal Micro Kit can be used directy in numerous downstream applications with no additional clean-up or purification required.  Downstream applications include: In vivo and in vitro introduction into cells and organisms Monoclonal and polyclonal antibody generation Mass spectrometry Gel electrophoresis Immunoassays Western blots

Figure 1. Endotoxin-Free Proteins. Endotoxins were removed from 100 µg protein samples in triplicate using the ProteoSpin™ Endotoxin Removal Micro Kit.  Samples containing 100 µg of BSA were spiked with endotoxins (0.9 EU/mg). The endotoxin-spiked BSA was then cleaned, in triplicate, using the ProteoSpinTM Endotoxin Removal Micro Kit by using 5 µL of the provided Endotoxin Removal Solution in the presence of Isopropanol. On average, the endotoxin levels in the 100 µg samples were reduced from 0.9 EU/mg to 0.005 EU/mg.

Figure 2. High Protein Recovery.   An average protein recovery of greater than 95% was achieved when endotoxins were removed from 100 µg inputs of protein in triplicate. Samples containing 100 µg of BSA were spiked with endotoxins (0.9 EU/mg). The endotoxin-spiked BSA was then cleaned, in triplicate, using the ProteoSpinTM Endotoxin Removal Micro Kit by using 5 µL of the provided Endotoxin Removal Solution in the presence of ethanol.  The BioRad Protein Assay was used to determine the amount of proteins present in the elutions, and 91 µg, 99 µg and 89 µg of BSA were recovered after the removal of the spiked endotoxins. There is very little loss of protein associated with using the ProteoSpin™ Endotoxin Removal Micro Kit.

Kit Specifications Maximum Protein Input 150 µg Maximum Protein Volume Input 450 µL Final Endotoxin Levels < = 0.01 EU/mg protein Average Recovery >95 % Minimum Elution Volume 50 µL Time to Complete 10 Purifications 20 minutes

ProteoSpin Endotoxin Removal Micro Kit Contents 1. Binding Buffer 2. Activation and Wash Buffer 3. Elution Buffer 4. Endotoxin Removal Solution 5. Neutralizer 6. Micro Spin Columns (20) 7. Elution Tubes (20) 8. Product Insert

Long-Term Storage: All solutions should be kept tightly sealed and stored at room temperature. All the reagents should remain stable for at least 2 years in their unopened containers.

Product Usage: For laboratory research use only, not for diagnostic use! Not for use in humans.

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