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Are CUSABIO recombinant proteins sterile?
Yes, we can offer an aseptic processing service and it is free of charge, but you should remark this information when placing the order. We've performed aseptic processing for liquid protein before lyophilization, but there may exist contamination during lyophilization process. To clarify, lyophilized proteins can’t guarantee aseptic processing for the whole manufacture process.
Can you remove the endotoxin?
Not all endotoxin can be removed. Please contact us in advance if you need to remove the tag which takes 2-3 business days. We could offer endotoxin removal service free of charge using PMB affinity chromatography, use LAL reagent to semi-quantitatively detect the content of endotoxin, and guarantee endotoxin level within 0.1 ng/μg (1 EU/μg).
How to avoid inclusion bodies and improve soluble expression?
a. For proteins with high hydrophobicity or transmembrane domains, you should add fusion tags or add heat shock chaperones, and induce for a shorter time at low temperature or change to poor media. Generate truncated forms of protein or use membrane-rich strains.
b. For incorrect disulfide bond formation, you should add fusion partners, including thioredoxin, DsbA, DsbC. Clone in a vector containing secretion signal peptide to cell periplasm. Use gamiB (DE3)strains with the oxidative cytoplasmic environment. Lower inducer concentration and induction temperature.
c. For incorrect folding, you should use a fusion partner. Co-express with molecular chaperones. Use strains with cold-adapted chaperones. Supplement media with chemical chaperones and cofactors. Reduce the inducer concentration and add fresh media. Induce for a shorter time at low temperature.