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In response to your enquiry, here is what Autogen Bioclear wrote: "The beta-defensin genes were directly expressed (no tags or leader sequence). Sequences are available (let me know if you require them). After fermentation, centrifugation, cell breakage; the proteins are folded into its active form during the folding/oxidation stage, this is when the disulfide bridges form. Specific processes are proprietary. The beta-defensins have a characteristic 6 cysteine structural motif containing 3 disulfide-bridges." I have tested the performance of your antibody against the denatured forms of my standards, HBD-1 (36aa and 47aa), HBD-2 and HBD-3. Unfortunately these results were negative and the only positive results were the native forms of HBD-1 36 and 47 aa, which gave a clear signal on the dot blot. Best regards
Asked on Nov 03 2005
Thank you for getting back to me with those details, it was very interesting. Unfortunately given that the source of the antisera have not been able to provide me with further details as to the synthesis of the immunising peptides it is difficult for me to comment on whether they have been raised against a correctly folded immunogen. If this was indeed the case then one might speculate that these antibodies only recognise the synthetic peptide and not the native form of the protein. However, checking back to past orders and this antibody is relatively popular and we have not received any complaints as such. I can but speculate that the absence of western or dot blot as applications on the datasheets is a reflection that these antibodies do not recognise their epitopes when immobilised on a membrane.
Answered on Nov 03 2005