Removal of endotoxin is one of the most difficult downstream processes during protein purification. EndoTrap® blue, as part of the EndoClear kit, is developed to meet the most challenging requirements of endotoxin removal: the highest endotoxin removal capacity combined with excellent recovery rates. EndoTrap® blue is based on affinity chromatography for column or batch mode. The working range of EndoTrap® blue is pH 4-9 and it works with an ionic strength up to 600 mM NaCl. Proteins, peptides, antibodies and plasmid DNA can be applied onto the EndoTrap® blue column. Customer specific buffers can be used when enriched freshly with Ca2+.
Besides the EndoTrap® blue, EndoTrap® red is also available. EndoTrap® red has a pH range of 6-9 and is capable to work with an ionic strength up to 250 mM ( <100 mM is recommended). LPS from Klebsiella pneumoniae and Serratia marcescens can only be removed with EndoTrap® red.
The detection of endotoxin is based on studies of Frederick Bang. He observed that bacteria caused intravascular coagulation in the American horseshoe crab, Limulus polyphemus. In collaboration, Levin and Bang found that the agent responsible for the clotting phenomena resided in the crab's amoebocytes, or circulating blood cells, and that pyrogen triggers the turbidity and gel-forming reaction enzymatically.
The Mini-LAL assay for endotoxin detection, the second part of the EndoClear kit, makes use of an activated enzyme. In the presence of a colorless substrate, the enzymatic reaction will cause a yellow colour to develop upon cleavage of the chromophore, p-nitroaniline (pNA). The reaction is stopped by the addition of acetic acid and the absorbance at 405 nm is measured with a spectrophotometer. The endotoxin concentration of samples with unknown concentrations, which are run concurrently with the standards, can be determined from a standard curveThe Profos EndoTrap® blue assay is intended for the removal of endotoxin with removal rates of >95% per cycle. The binding capacity of EndoTrap® blue is 2,000,000 EU/ml (1EU = 100 pg LPS)
The Hycult Biotech Mini-LAL assay is intended for the quantitative detection of endotoxin in culture medium, buffers, and other solutions. The kit has a minimum endotoxin detection limit of 0.01 EU/ml and a measurable concentration range of 0.01 to 10 EU/ml.- Binding capacity of 2,000,000 EU/ml.
- Endotoxin removal rates of >95% per cycle.
- Final LPS concentration 0.005 - 0.1 EU/ml.
- Column or batch mode.
- Operating at pH 4.0-9.0.
- Suitable for plasmid DNA, proteins, peptides and antibodies.
- Suitable for an ionic strength up to 600 mM NaCl.
- Customer specific buffers can be used when enriched freshly with Ca2+.
Hycult Biotech Mini-LAL assay (endotoxin detection):
- Minimum concentration which can be measured is 0.01 EU/ml.
- Measurable endotoxin detection range of 0.01-10 EU/ml.
- Working volume of 50 µl/well.
- 72 determinations
- Operating at pH 6.5-8.0.
- The Profos EndoTrap® blue Assay is based on affinity chromatography with a working time of ~30 minutes.
- Sample is applied to the column with equilibration buffer.
- Fractions are collected.
- Endotoxin removal rate is >95% per cycle.
- Samples are measured with Hycult Biotech LAL Assay.
Hycult Biotech Mini-LAL assay (endotoxin detection):
- The LAL assay is an enzyme-based assay with a working time of ~45 minutes.
- Samples and standards are incubated with LAL reagent.
- The enzymatic reaction, triggered by endotoxin, will cause a yellow color to develop upon cleavage of the chromophore, p-nitroaniline.
- The enzymatic reaction is stopped by the addition of acetic acid.
- The absorbance at 405 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the E. coli standards (log).
- The endotoxin concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
The Profos EndoTrap® blue columns should be stored in regeneration buffer with 0.02% sodium azide.
After reconstitution the Hycult Biotech Mini-LAL assay reagents (E. coli standard and LAL reagent) should be stored in aliquots at -20 °C.- Gonçalves, S et al; Associations between renal function, volume status and endotoxaemia in chronic kidney disease patients. Nephrol Dial Transpl 2006, 21: 2788
- Bowdish, D et al; The human cationic peptide LL-37 induces activation of the extracellular signal-regulated kinase and p38 kinase pathways in primary human monocytes. J Immunol 2004, 172: 3758
- Bublin, M et al; Use of a genetic cholera toxin B subunit/allergen fusion molecule as mucosal delivery system with immunosuppressive activity against Th2 immune responses. Vaccine 2007, 25: 8395
- Cordery, D et al; Characterization of a Plasmodium falciparum macrophage-migration inhibitory factor homologue. J Infect Dis 2007, 195: 905
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use or derivation of this product.
All materials coming in contact with specimen or test material should be endotoxin-free.- Gonçalves, S et al; Associations between renal function, volume status and endotoxaemia in chronic kidney disease patients. Nephrol Dial Transpl 2006, 21: 2788
- Bowdish, D et al; The human cationic peptide LL-37 induces activation of the extracellular signal-regulated kinase and p38 kinase pathways in primary human monocytes. J Immunol 2004, 172: 3758
- Bublin, M et al; Use of a genetic cholera toxin B subunit/allergen fusion molecule as mucosal delivery system with immunosuppressive activity against Th2 immune responses. Vaccine 2007, 25: 8395
- Cordery, D et al; Characterization of a Plasmodium falciparum macrophage-migration inhibitory factor homologue. J Infect Dis 2007, 195: 905