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Dear Sir or Madam, My name is XXXX, and I am a biologist of a biotech company called STAT-Diagnostica, which is located in the Parc Científic de Barcelona (Spain). I am interested in performing a Direct Sandwich ELISA assay and I would like to consult some questions regarding the selection of antibodies and human troponin protein. I have seen that you recommend anti-cardiac troponin I antibody [284 (19C7)]-ref. ab10236 due to their high sensitivity, good kinetics, low background and high reproducibility. I would use it as capture antibody: - could you please tell me what is the recommended concentration for coating the well plates?, and also - what would be the optimum pH of the coating buffer for this antibody, 9,6 or 7.4? On the other hand, detection antibodies could be anti-cardiac troponin I antibody [MF4]-HRP-ref.ab10239 and anti-cardiac troponin antibody [284(19C7)]-ref.ab19615. Is it correct? Could you tell me what would approximately be the optimal assay concentration? Furthermore, I would like to use Cardiac Troponin I protein ref. ab9936 as a standard, but I see that It has been only tested for WB. Do you have information about using it for ELISA? In other case, do you have a specific one for ELISA? Lastly, I would like to know what would be the delivery time. I would appreciate your comments. Many thanks in advance. Best Regards,
Asked on Sep 07 2011
Thank you for your email. We have updated the datasheet of ab10236; few of the match pairs are no longer available in our catalogue so we have removed these from the datasheet. We recommend using ab10236 as a capture ab along with ab10239 as a detection antibody. In general many general protocols could be used with our cTnI antibodies and the optimal working concentrations should be determined separately for each application by the user. In our own lab we are using a special two-step immunofluorometric assay and streptavidin-coated plates with biotinylated capture MAbs and detection MAbs which are conjugated with stable europium chelate. The MAb concentrations in this kind of assay are different than what are needed in some other kind of assay. With very general protocol using PBS buffers and HRP detection method I can only recommend to start testing with something like 5 ug/ml MAb concentration for capture MAb and 0.5-1 ug/ml concentration for HRP-conjugated detection MAb and then to start optimization from there. Regarding pH I would recommend 7.4. I hope that this information is helpful to you. Should you have any question please do not hesitate to contact me.
Answered on Sep 07 2011