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Please find attached the QC Data sheets for the secondary antibodies that were used in the experiment. I have called Thermo Scientific/Pierce’s technical department and they recommended using product no. 31462 because it was cross absorbed and would give no cross reactivity. As described in previous emails, the mouse monoclonal antibody (ab14422) was able be detected by an anti-rabbit secondary antibody. An anti-mouse antibody was used as a control. The experiment was repeated a second time, and still produced the same results. I later informed you that the antibodies that were purchased are widely commercialized and we have had no problems with them cross reacting with other species. We have used these antibodies in other assays as well. Due to confidentiality purposes, we still can not release the information about the concentration of the peptide or the concentration of the antibodies used. Therefore, I would like to request a replacement for this product or a refund.
Asked on Nov 09 2011
Thank you for the datasheets of the secondary antibodies. I am surprised that Thermo recommended 31462. The datasheet for this secondary includes this note: The product has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with human serum proteins. However, this antibody may cross-react with immunoglobulins from other species. This is in the "Antibody Specificity" field of the datasheet. You may want to ask them about that, because it suggests that reactivity with mouse IgG was not tested. I will find out from the laboratory that produced the antibody how it was validated for ELISA and will forward this information to you. If you can confirm that your protocol is similar, with regards to the concentration of peptide and antibody, and the protocol, I will be happy to send a replacement or refund. I assure you I want to resolve this quickly, hopefully before the end of the week.
Answered on Nov 09 2011