Endotoxin Removal Kit II
First Edition (Revised on April, 2016)
Bacterial endotoxin is the unique component of gram negative bacteria cell wall. It’s the exogenous pyrogen which could activate macrophages, neutrophils, etc. to release Inflammatory mediators, and further to cause injury of the body, septic shock or even death. As endotoxin is an important pollutant of biological products, it is crucial to strictly control endotoxin level in biological products and medicines, which have direct access to human and animals. Therefore, the removal of endotoxin is very important in downstream purification process of biological products.
Extract method is used for removal of endotoxin in this kit, based on the hydrophobicity of lipopolysaccharide of bacterial endotoxin.
REAGENTS AND MATERIALS PROVIDED
STORAGE AND PERIOD OF VALIDITY
Store at room temperature. The Period of Validity is 1 year.
Endotoxin removal of hydrosoluble proteins, peptides, antibodies, etc. The final Endotoxin level would be less than 0.1 EU/ml.
1. Prepare endotoxin free centrifugal tubes, trips, receiving tubes etc.
2. Add reagent A into samples with a final concentration of 1-2% (V/V). Put the mixed samples in ice bath with stirring for 5-15min.
3. Water bath at 50-75℃ for 1-5min. If the protein is not stable, please water bath at 37℃ for 5-10min. The liquid will be layered.
4. Centrifuge at 12000g, 30℃ for 1min. Then, collect the supernates for later use. If the concentration of endotoxin does not meet the need, repeat step 2-3.
1. Please strictly control the operating environment of the experiment to avoid outside endotoxin.
2. For your safety and health, please wear a lab-gown and disposable gloves in experiments.