Generating Custom Antibodies against a Gel Band Antigen
When Gary approached us to develop an antibody against his expressed protein, he was concerned about the insolubility of his protein and the presence of non-specific proteins in his sample.
We discussed how it’s okay for protein antigens to be insoluble and why this won’t negatively affect the ability of the antigen to elicit an immune response. But, we also recommended that he consider running the protein on a gel and cutting out the band that corresponded to his protein.
We explained how using a gel band antigen such as this is highly effective for purifying and isolating the target protein. And, we assured him that gel bands work well as antigens without any further modification necessary. Gary followed the guidelines discussed on our Gel Band Proteins page and sent everything to us following the steps listed on our How to Prepare your Protein page.
John sent over the cut out gel bands the following week and we began immunizations immediately. Seven weeks later, we shipped the first production bleeds to Gary so that he could test reactivity against the native protein. Two days later, he sent us an email expressing how excited he was to already see a strong signal against the target protein on a Western Blot. As the project continued and came to a close, he confirmed that antibody sensitivity against the protein seemed to increase as the affinity maturation process completed.
Nine months after the project was completed, Gary sent a grateful email with a link to a research paper that he and his colleagues had just published.