The DNA fragment corresponding to Leu29-Gly245 of the human CD276 antigen is fused with an FC-Myc-tag at the C-terminus and then is expressed in the mammalian cells. The resulting product is the recombinant human CD276 antigen. Its purity is over 90% measured by SDS-PAGE. On the gel, this protein was migrated to the molecular weight band of about 60-80 kDa. The endotoxin content of this CD276 protein is less than 1.0 EU/ug as determined by the LAL method. Its bioactivity has been validated through an ELISA. In the functional ELISA, this recombinant CD276 protein can bind to the anti-CD276 antibody, and the EC50 is 1.961-2.243 ng/ml. This CD276 protein is available now.
CD276 is an immune checkpoint molecule in the epithelial-mesenchymal transition (EMT) pathway. It is involved in cell proliferation, invasion, and metastasis in malignancies. Overexpression of CD276 has been found in numerous solid tumors such as breast, colorectal, and lung cancer, and has been strongly linked to a poor clinical prognosis.
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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.
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